Cyclic peptides multimers targeting alpha-4-beta-7 integrin

ABSTRACT

There is described herein, multimers comprising a plurality of compounds covalently linked together, the compounds independently being of formula (I).

RELATED APPLICATION

This application is a national phase application under 35 U.S.C. § 371 of International Application No. PCT/CA2017/000244 filed Nov. 10, 2017, which claims priority to U.S. Provisional Application No. 62/421,117, filed on Nov. 11, 2016, both of which are incorporated herein in their entirety.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Dec. 20, 2019, is named 50412-120002_Sequence_Listing_12.19.19_ST25 and is 112,362 bytes in size.

FIELD OF THE INVENTION

The invention relates to antagonists of α4β7 integrin, and more particularly to cyclic peptide antagonists.

BACKGROUND OF THE INVENTION

Integrins are transmembrane receptors that are the bridges for cell-cell and cell-extracellular matrix (ECM) interactions. When triggered, integrins trigger chemical pathways to the interior (signal transduction), such as the chemical composition and mechanical status of the ECM.

Integrins are obligate heterodimers, having two different chains: the α (alpha) and β (beta) subunits.

The α4β7 integrin is expressed on lymphocytes and is responsible for T-cell homing into gut-associated lymphoid tissues through its binding to mucosal addressin cell adhesion molecule (MAdCAM), which is present on high endothelial venules of mucosal lymphoid organs. Inhibitors of specific integrin-ligand interactions have been shown effective as anti-inflammatory agents for the treatment of various autoimmune diseases. For example, monoclonal antibodies displaying high binding affinity for α4β7 have displayed therapeutic benefits for gastrointestinal auto-inflammatory/autoimmune diseases, such as Crohn's disease, and ulcerative colitis.

There is a need to develop improved α4β7 antagonists to prevent or treat inflammatory conditions and/or autoimmune diseases.

Certain methods of making cyclic peptides (nacellins) are described in Applicant's PCT Publication No. WO 2010/105363.

SUMMARY OF THE INVENTION

In an aspect, there is provided, a multimer comprising a plurality of compounds covalently linked together, the compounds independently being of formula (I):

wherein

-   R¹ is H; lower alkyl; aryl; heteroaryl; alkenyl; or heterocycle; all     of which are optionally substituted at one or more substitutable     positions with one or more suitable substituents; -   R² and R³ are each independently an amino acid chain of a     proteinogenic or a non-proteinogenic alpha-amino acid,     -   provided that R² and R³ may be covalently linked to each other         to form a ring; -   R⁴ and R⁵ are each independently H; lower alkyl; aryl; heteroaryl;     alkenyl; heterocycle; acids of the formula —C(O)OH; esters of the     formula —C(O)OR* wherein R* is selected from alkyl and aryl; amides     of the formula —C(O)NR**R***, wherein R** and R*** are independently     selected from H, alkyl and aryl; —CH₂C(O)R, wherein R is selected     from —OH, lower alkyl, aryl, -loweralkyl-aryl, or —NRaRb, where Ra     and Rb are independently selected from H, lower alkyl, aryl or     -loweralkyl-aryl; or —C(O)Rc, wherein Rc is selected from lower     alkyl, aryl or -lower alkyl-aryl; or -lower alkyl-ORd, wherein Rd is     a suitable protecting group or OH group; all of which are optionally     substituted at one or more substitutable positions with one or more     suitable substituents;     -   provided that R² or R³ can be covalently linked to R¹ to form a         cyclic secondary amine, and/or to R⁴ or R⁵ to form a ring, R⁴         and R⁵ may also be covalently linked to each other to form a         ring; -   R⁶ is H, lower alkyl, benzyl, alkenyl, lower alkyloxy; aryl;     heteroaryl; heterocycle; —C(O)R****, wherein R**** is independently     selected from alkyl, aryl, heteroaryl, amino, aminoalkyl, aminoaryl,     aminoheteroaryl, alkoxy, aryloxy, heteroaryloxy; —CH₂C(O)R; or     —C(O)Rc; all of which are optionally substituted at one or more     substitutable positions with one or more suitable substituents,     -   or along with R⁷ or R⁸, a cyclic side chain of a proteinogenic         or a non-proteinogenic amino acid having, the N-terminus thereof         being the N—R⁶, wherein the proteinogenic or a non-proteinogenic         amino acid can be substituted with a suitable substituent; -   R⁷ and R⁸ are independently selected from the amino acid side chains     of a proteinogenic or a non-proteinogenic alpha-amino acid having     the N-terminus thereof being the N—R⁶, or may form a cyclic side     chain with R⁶; -   stereocentres 1*, 2* and 3* are each independently selected from R     and S; -   n is 1, 2, 3, or 4 and where n is 2-4, each R⁷ and each R⁸ are     independent of each other; and -   wherein Z is an amino terminus of an amino acid; —C═O— adjacent L is     the carboxy terminus of an amino acid; and L along with Z and —C═O—     is a peptide having the following formula:     X^(y)—X^(z)—X¹—X²—X³     -   wherein X^(y) and X^(z) are each independently a proteinogenic         or non-proteinogenic amino acid;     -   X¹ is Leucine or tert-butyl-Ala;     -   X² is Asp; and     -   X³ is any amino acid listed under column X³ of Table 1B.

In an aspect, there is provided, a pharmaceutical composition comprising the multimer described herein along with the pharmaceutically acceptable carrier. The pharmaceutical composition may be formulated for any one of oral delivery, topical delivery and parenteral delivery.

In an aspect, there is provided, a method of treating inflammation or an autoimmune disease in a patient, comprising administering to the patient a therapeutically effective amount of the multimer described herein. Preferably the inflammation or an autoimmune disease is gastrointestinal.

In an aspect, there is provided, a method for treating a condition in a patient associated with a biological function of an α4β7 integrin, the method comprising administering to the patient a therapeutically effective amount of the multimer described herein.

In an aspect, there is provided, a method for treating a disease or condition in a patient comprising administering to the patient a therapeutically effective amount of the multimer described herein, wherein the disease or condition is a local or systemic infection of a virus or retrovirus.

In an aspect, there is provided, a method for treating a disease or condition in a patient comprising administering to the patient a therapeutically effective amount of the multimer described herein, wherein the hepatitis A, B or C, hepatic encephalopathy, non-alcoholic steatohepatitis, cirrhosis, variceal bleeding, hemochromatosis, Wilson disease, tyrosinemia, alpha-1-antitrypsin deficiency, glycogen storage disease, hepatocellular carcinoma, liver cancer, primary biliary cholangitis, primary sclerosing cholangitis, primary biliary sclerosis, biliary tract disease, autoimmune hepatitis, or graft-versus-host disease.

BRIEF DESCRIPTION OF FIGURES AND TABLES

These and other features of the preferred embodiments of the invention will become more apparent in the following detailed description in which reference is made to the appended drawings and tables wherein:

FIG. 1 shows representative compounds of the present application, namely from the following classes, 18-membered ring, 21-membered ring, 21-membered ring (non-canonical, i.e. having a delta amino acid), 22-membered ring, and 24-membered ring.

FIG. 2 shows a representative 18-membered ring compound along with variations made at certain positions with corresponding α4β7 integrin ELISA IC50 binding values associated with those variations.

FIG. 3 shows a representative 21-membered ring compound along with variations made at certain positions with corresponding α4β7 integrin ELISA IC50 binding values associated with those variations.

FIG. 4 shows a representative 21-membered ring (non-canonical, i.e. having a delta amino acid) compound along with variations made at certain positions with corresponding α4β7 integrin ELISA IC50 binding values associated with those variations.

FIG. 5 shows a representative 22-membered ring compound along with variations made at certain positions with corresponding α4β7 integrin ELISA IC50 binding values associated with those variations.

FIGS. 6A and 6B show representative NMR data for a multimeric molecule, Compound No. 390, with ¹H— and ¹H—¹H TOCSY NMR spectra recorded at 25° C.

FIG. 7 shows the binding to α4β7 integrin measured as a MADCAM-1 competition assay in human whole blood for: a) representative monomeric Compound 456 (ET4062) and multimeric Compound No.s 534 (ET4113) and 535 (ET4110), derived from Compound 456, and; b) representative monomeric Compound 340 (ET2451) and multimeric Compound No.s 390 (ET3755) and 517 (ET3764), derived from Compound 340.

FIG. 8 Shows the detection of α4β7+ Th memory cells trafficking in the mesenteric lymph nodes in mice suffering from DSS-induced colitis treated for 4 days with Compound No. 517 (ET3764) or vehicle.

FIG. 9 shows the α4β7+ Th memory lymphocyte content in mesenteric lymph nodes taken from mice exposed to DSS irritant and treated for 4 days with various concentrations of Compound No. 517 (ET3764) or control (SMEDDS vehicle)

FIG. 10 shows the receptor occupancy of representative multimeric compounds on α4β7-positive T helper memory cells as measured in a MADCAM-1 competition assay in human whole blood.

FIG. 11 shows the receptor occupancy of representative nacellin dimers on α4β7-negative Th memory cells as measured in a VCAM-1 competition assay in human whole blood.

Table 1 shows compounds exhibiting α4β7 integrin affinity, selectivity and/or activity; and specifically with respect to these compounds: (A) the structure of the linker portion; (B) the structure of the peptide portion; and (C) and (C′) the affinity, selectivity and activity values.

To aid reading of the table, the following is noted:

Table 1A:

-   -   If R2 is H and R3 is CH3, the carbon atom bearing R2 and R3 has         S-configuration.     -   If R2 is CH3 and R3 is H, the carbon atom bearing R2 and R3 has         R-configuration.     -   If R2 is H and R3 is CH2-S-Ph, the carbon atom bearing R2 and R3         has S-configuration.     -   If R4 is H and R5 is C(O)—NH-tert-Butyl, the carbon atom bearing         R4 and R5 has S-configuration.     -   If R4 is C(O)—NH-tert-Butyl and R5 is H, the carbon atom bearing         R4 and R5 has R-configuration.     -   If R1 and R2 are both Pro-, the R1 and R2 substituents are         covalently bound and form the pyrrolidine ring of Pro.

Table 1B

-   -   If R6 and R7 are both Pro, the R6 and R7 substituents are         covalently bound and form the pyrrolidine ring of Pro.     -   If R6 and R8 are both dPro, the R6 and R8 substituents are         covalently bound and form the pyrrolidine ring of dPro.     -   If R6 and R7 are both [(4S)-fluoro-Pro], the R6 and R7         substituents are covalently bound and form the pyrrolidine ring         of [(4S)-fluoro-Pro].     -   If R7 is Nva and R8 is H, the carbon atom bearing R7 and R8 has         S-configuration.     -   If R6 and R7 are both Hyp, the R6 and R7 substituents are         covalently bound and form the pyrrolidine ring of Hyp.     -   If no entry exists under column Xz, the residue is absent.

Table 1C and 1C′

-   -   If no entry exists under any of the columns, no data was         collected.

Table 1X is a correspondence table linking the compounds described herein with the synthesis protocols outlined in the methods and materials.

Table 2 shows multimeric compounds exhibiting α4β7 integrin affinity, selectivity and/or activity; and specifically with respect to these compounds: (A) the structure of the linker portion; (B) the structure of the peptide portion; and (C) the affinity, selectivity and activity values.

To aid reading of the table, the following is noted:

Table 2A

-   -   If R2 is H and R3 is CH3, the carbon atom bearing R2 and R3 has         S-configuration.

Table 2B

-   -   If R6 and R7 are both Pro, the R6 and R7 substituents are         covalently bound and form the pyrrolidine ring of Pro.     -   If R6 and R7 are both Hyp, the R6 and R7 substituents are         covalently bound and form the pyrrolidine ring of Hyp.     -   If no entry exists under column Xz, the residue is absent.

Table 2X is a correspondence table linking the multimers described herein with the synthesis protocols outlined in the methods and materials. m/z is (M+2H/2) and additional information regarding the linker.

Table 3 is a table of the sequence listing

DETAILED DESCRIPTION

In the following description, numerous specific details are set forth to provide a thorough understanding of the invention. However, it is understood that the invention may be practiced without these specific details.

In an aspect, there is provided, a multimer comprising a plurality of compounds covalently linked together, the compounds independently being of formula (I):

wherein

R¹ is H; lower alkyl; aryl; heteroaryl; alkenyl; or heterocycle; all of which are optionally substituted at one or more substitutable positions with one or more suitable substituents;

R² and R³ are each independently an amino acid chain of a proteinogenic or a non-proteinogenic alpha-amino acid,

-   -   provided that R² and R³ may be covalently linked to each other         to form a ring;

R⁴ and R⁵ are each independently H; lower alkyl; aryl; heteroaryl; alkenyl; heterocycle; acids of the formula —C(O)OH; esters of the formula —C(O)OR* wherein R* is selected from alkyl and aryl; amides of the formula —C(O)NR**R***, wherein R** and R*** are independently selected from H, alkyl and aryl; —CH₂C(O)R, wherein R is selected from —OH, lower alkyl, aryl, -loweralkyl-aryl, or —NRaRb, where Ra and Rb are independently selected from H, lower alkyl, aryl or -loweralkyl-aryl; or —C(O)Rc, wherein Rc is selected from lower alkyl, aryl or -lower alkyl-aryl; or -lower alkyl-ORd, wherein Rd is a suitable protecting group or OH group; all of which are optionally substituted at one or more substitutable positions with one or more suitable substituents;

-   -   provided that R² or R³ can be covalently linked to R¹ to form a         cyclic secondary amine, and/or to R⁴ or R⁵ to form a ring, R⁴         and R⁵ may also be covalently linked to each other to form a         ring;

R⁶ is H, lower alkyl, benzyl, alkenyl, lower alkyloxy; aryl; heteroaryl; heterocycle; —C(O)R****, wherein R**** is independently selected from alkyl, aryl, heteroaryl, amino, aminoalkyl, aminoaryl, aminoheteroaryl, alkoxy, aryloxy, heteroaryloxy; —CH₂C(O)R; or —C(O)Rc; all of which are optionally substituted at one or more substitutable positions with one or more suitable substituents,

-   -   or along with R⁷ or R⁸, a cyclic side chain of a proteinogenic         or a non-proteinogenic amino acid having, the N-terminus thereof         being the N—R⁶, wherein the proteinogenic or a non-proteinogenic         amino acid can be substituted with a suitable substituent;

R⁷ and R⁸ are independently selected from the amino acid side chains of a proteinogenic or a non-proteinogenic alpha-amino acid having the N-terminus thereof being the N—R⁶, or may form a cyclic side chain with R⁶;

stereocentres 1*, 2* and 3* are each independently selected from R and S;

n is 1, 2, 3, or 4 and where n is 2-4, each R⁷ and each R⁸ are independent of each other; and

wherein Z is an amino terminus of an amino acid; —C═O— adjacent L is the carboxy terminus of an amino acid; and L along with Z and —C═O— is a peptide having the following formula: X^(y)—X^(z)—X¹—X²—X³

-   -   wherein X^(y) and X^(z) are each independently a proteinogenic         or non-proteinogenic amino acid;     -   X¹ is Leucine or tert-butyl-Ala;     -   X² is Asp; and     -   X³ is any amino acid listed under column X³ of Table 1B.

The compounds shown in Tables 1A, 1B and 1C (and 1C′) exhibit antagonistic activity against α4β7 integrin and having selectivity over α4β1 integrin. A person skilled in the art would expect that substituents R¹-R⁸ and amino acids X^(y), X^(z), X¹, X², and X³ outlined in Tables 1A and 1B with respect to different compounds could be combined in any manner and would likely result in a compound that would exhibit α4β7 integrin activity and selectivity. These compounds are further described in WO 2017/079820, the entirety of which is incorporated herein by reference.

Multimerized, specifically dimerized, versions of certain compounds described herein exhibited affinity, selectivity and activity, summarized in Tables 2A, 2B and 2C.

As used herein, the term “amino acid” refers to molecules containing an amine group, a carboxylic acid group and a side chain that varies. Amino acid is meant to include not only the twenty amino acids commonly found in proteins but also non-standard amino acids and unnatural amino acid derivatives known to those of skill in the art, and therefore includes, but is not limited to, alpha, beta and gamma amino acids. Peptides are polymers of at least two amino acids and may include standard, non-standard, and unnatural amino acids. A peptide is a polymer of two or more amino acids.

The following abbreviations are used herein:

Abbreviation Description 1,2-cis-ACHC cis-2-aminocyclohexanecarboxylic acid 1,2-trans-ACHC trans-2-aminocyclohexanecarboxylic acid 1Nal 1-napthylalanine 2Abz anthranilic acid, 2-aminobenzoic acid 2lgl 2-indanylglycine 2Nal 2-napthylalanine Abu 2-aminobutyric acid Aic aminoindan-2-carboxylic acid allolle allo-sioleucine, (2S,3R)-2-amino-3-methylpentanoic acid alloThr allo-threonine, (2S,3S)-2-amino-3-hydroxybutyric acid alphaMePhe α-methyl-phenylalanine, (S)-(−)-2-amino-2-methyl-3-phenylpropionic acid Asp(ethyl ester) aspartic acid β-ethyl ester Atc 2-aminotetraline-2-carboxylic acid Aze azetidine-2-carboxylic acid BHT butylated hydroxytoluene Bip biphenylalanine C10 sebacic acid C12 dodecanedioic C7 pimelic acid C8 suberic acid C9 azelaic acid Cha β-cyclohexyl alanine, (S)-2-amino-3-cyclohexylpropionic acid Chg cyclohexyl glycine cis-dhyp cis-D-4-Hydroxyproline, (2R,4R)-4-Hydroxypyrrolidine-2-carboxylic acid cycloLeu cyclo leucine, 1-Aminocyclopentane-1-carboxylic acid cyclopropylAla β-cyclopropyl alanine, (S)-2-amino-3-cyclopropyl-propionic acid d2lgl 2-indanyl-D-glycine Dap(Cbz) Nβ-2-2,3-diaminopropionic acid DBU 1,8-diazabicyclo[5.4.0]undec-7-ene DEPBT 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one dHyp trans-D-4-hydroxyproline, (2R,4S)-4-hydroxypyrrolidine-2-carboxylic acid DIAD diisopropyl azodicarboxylate DIG diglycolic acid DIPEA N,N-diisopropylethylamine DMAP 4-(Dimethylamino)pyridine dMeArg N-methyl-D-arginine dMebetaHomoLys N-methyl-D-β-homoLys dMeLys N-methyl-D-Lysine DMF N,N-dimethylformamide DMSO dimethyl sulfoxide dNle D-norleucine dOrn D-ornithine dOrn(dimethyl) Nδ-dimethyl-D-ornithine dPip D-pipecolic acid, D-homoPro dSer(OBn) O-benzyl-D-serine dTic (3R)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid dTiq D-1,2,3,4-tetrahydroisoquinoline-1-carboxylic acid dTyr(OAllyl) O-allyl-D-tyrosine dTyr(OBn) O-benzyl-D-tyrosine EDC N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride Fmoc 9-fluorenylmethoxycarbonyl HATU O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate HCTU 2-(6-chloro-1H-benzotriazole-1-yl)-1,1,3,3-tetramethylaminium hexafluorophosphate HFIP 1,1,1,3,3,3-hexafluoro-2-propanol His(Bn) Nτ-benzyl-histidine HomocycloLeu homocyclo leucine, 1-Aminocyclohexanecarboxylic acid Hyp trans-4-hydroxyproline, (2S,4R)-4-hydroxypyrrolidine-2-carboxylic acid Hyp(OBn) O-benzyl-trans-4-hydroxyproline MeAsp N-methyl aspartic acid MebetaHomoLys N-methyl β-homoLysine MebetaHomoLys(Me)2 Nα-methyl-Nε-dimethyl-β-homoLysine MeLeu N-methyl leucine MeMet N-methyl methionine MePhe N-methyl phenylalanine metaY(Opr) metaTyrosine MeThr N-methyl threonine MeTyr N-methyl tyrosine NMP N-methylpyrrolidone Nosyl chloride 2-nitrobenzenesulfonyl chloride Nva norvaline Orn(acetamide) Nδ-acetamide-ornithine Orn(benzamide) Nδ-benzamide-ornithine Orn(ethylcarbamate) Nδ-ethylcarbamate-ornithine Orn(methanesulfonamide) Nδ-methanesulfonamide-ornithine Orn(pentyl amide) Nδ-pentyl amide-ornithine PDA 1,4-phenyldiacetic acid Pen penicillamine, β,β-dimethyl-cysteine Pip pipecolic acid, homoPro Sar sarcosine, N-methyl glycine tertbutylAla β-tert-butyl alanine, neopentylglycine TFA trifluoroacetic acid TFE 2,2,2-Trifluoroethanol THF tetrahydrofuran Thr(OBn) O-benzyl-threonine Thr(OEt) O-ethyl-threonine Thr(OMe) O-methyl-threonine Tic (3S)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid TIS triisopropylsilane Tyr(2-methoxy diaryl O-2-methoxy-phenyl-tyrosine ether) Tyr(2-tolyl diaryl ether) O-2-methyl-phenyl-tyrosine Tyr(3,4-difluoro diaryl O-3,4-difluoro-phenyl-tyrosine ether) Tyr(3,4-dimethyl diaryl O-3,4-dimethyl-phenyl-tyrosine ether) Tyr(3-CO2Me diaryl ether) O-3-methylester-phenyl-tyrosine Tyr(3-fluoro diaryl ether) O-3-fluoro-phenyl-tyrosine Tyr(3-methoxy diaryl O-3-methoxy-phenyl-tyrosine ether) Tyr(3-methyl diaryl ether) O-3-methyl-phenyl-tyrosine Tyr(4-CF3 diaryl ether) O-4-trifluoromethyl-phenyl-tyrosine Tyr(4-CO2H diaryl ether) O-4-carboxylate-phenyl-tyrosine Tyr(4-CO2Me diaryl ether) O-4-methylester-phenyl-tyrosine Tyr(4-fluoro diaryl ether) O-4-fluoro-phenyl-tyrosine Tyr(4-methoxy diaryl O-4-methoxy-phenyl-tyrosine ether) Tyr(OAllyl) O-allyl-tyrosine Tyr(OPh) O-phenyl-tyrosine vinyl-Br-Leu 2-amino-4-bromo-4-pentenoic acid

The term “suitable substituent” as used in the context of the present invention is meant to include independently H; hydroxyl; cyano; alkyl, such as lower alkyl, such as methyl, ethyl, propyl, n-butyl, t-butyl, hexyl and the like; alkoxy, such as lower alkoxy such as methoxy, ethoxy, and the like; aryloxy, such as phenoxy and the like; vinyl; alkenyl, such as hexenyl and the like; alkynyl; formyl; haloalkyl, such as lower haloalkyl which includes CF₃, CCl₃ and the like; halide; aryl, such as phenyl and napthyl; heteroaryl, such as thienyl and furanyl and the like; amide such as C(O)NR_(a)R_(b), where R_(a) and R_(b) are independently selected from lower alkyl, aryl or benzyl, and the like; acyl, such as C(O)—C₆H₅, and the like; ester such as —C(O)OCH₃ the like; ethers and thioethers, such as O-Bn and the like; thioalkoxy; phosphino; and —NR_(a)R_(b), where R_(a) and R_(b) are independently selected from lower alkyl, aryl or benzyl, and the like. It is to be understood that a suitable substituent as used in the context of the present invention is meant to denote a substituent that does not interfere with the formation of the desired product by the processes of the present invention.

As used in the context of the present invention, the term “lower alkyl” as used herein either alone or in combination with another substituent means acyclic, straight or branched chain alkyl substituent containing from one to six carbons and includes for example, methyl, ethyl, 1-methylethyl, 1-methylpropyl, 2-methylpropyl, and the like. A similar use of the term is to be understood for “lower alkoxy”, “lower thioalkyl”, “lower alkenyl” and the like in respect of the number of carbon atoms. For example, “lower alkoxy” as used herein includes methoxy, ethoxy, t-butoxy.

The term “alkyl” encompasses lower alkyl, and also includes alkyl groups having more than six carbon atoms, such as, for example, acyclic, straight or branched chain alkyl substituents having seven to ten carbon atoms.

The term “aryl” as used herein, either alone or in combination with another substituent, means an aromatic monocyclic system or an aromatic polycyclic system. For example, the term “aryl” includes a phenyl or a napthyl ring, and may also include larger aromatic polycyclic systems, such as fluorescent (eg. anthracene) or radioactive labels and their derivatives.

The term “heteroaryl” as used herein, either alone or in combination with another substituent means a 5, 6, or 7-membered unsaturated heterocycle containing from one to 4 heteroatoms selected from nitrogen, oxygen, and sulphur and which form an aromatic system. The term “heteroaryl” also includes a polycyclic aromatic system comprising a 5, 6, or 7-membered unsaturated heterocycle containing from one to 4 heteroatoms selected from nitrogen, oxygen, and sulphur.

The term “cycloalkyl” as used herein, either alone or in combination with another substituent, means a cycloalkyl substituent that includes for example, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.

The term “cycloalkyl-alkyl-” as used herein means an alkyl radical to which a cycloalkyl radical is directly linked; and includes, but is not limited to, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, 1-cyclopentylethyl, 2-cyclopentylethyl, cyclohexylmethyl, 1-cyclohexylethyl and 2-cyclohexylethyl. A similar use of the “alkyl” or “lower alkyl” terms is to be understood for aryl-alkyl-, aryl-loweralkyl- (eg. benzyl), -lower alkyl-alkenyl (eg. allyl), heteroaryl-alkyl-, and the like as used herein. For example, the term “aryl-alkyl-” means an alkyl radical, to which an aryl is bonded. Examples of aryl-alkyl- include, but are not limited to, benzyl (phenylmethyl), 1-phenylethyl, 2-phenylethyl and phenylpropyl.

As used herein, the term “heterocycle”, either alone or in combination with another radical, means a monovalent radical derived by removal of a hydrogen from a three- to seven-membered saturated or unsaturated (including aromatic) cyclic compound containing from one to four heteroatoms selected from nitrogen, oxygen and sulfur. Examples of such heterocycles include, but are not limited to, aziridine, epoxide, azetidine, pyrrolidine, tetrahydro-furan, thiazolidine, pyrrole, thiophene, hydantoin, diazepine, imidazole, isoxazole, thiazole, tetrazole, piperidine, piperazine, homopiperidine, homopiperazine, 1,4-dioxane, 4-morpholine, 4-thiomorpholine, pyridine, pyridine-N-oxide or pyrimidine, and the like.

The term “alkenyl”, as used herein, either alone or in combination with another radical, is intended to mean an unsaturated, acyclic straight chain radical containing two or more carbon atoms, at least two of which are bonded to each other by a double bond. Examples of such radicals include, but are not limited to, ethenyl (vinyl), 1-propenyl, 2-propenyl, and 1-butenyl.

The term “alkynyl”, as used herein is intended to mean an unsaturated, acyclic straight chain radical containing two or more carbon atoms, at least two of which are bonded to each other by a triple bond. Examples of such radicals include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, and 1-butynyl.

The term “alkoxy” as used herein, either alone or in combination with another radical, means the radical —O—(C_(1-n))alkyl wherein alkyl is as defined above containing 1 or more carbon atoms, and includes for example methoxy, ethoxy, propoxy, 1-methylethoxy, butoxy and 1,1-dimethylethoxy. Where n is 1 to 6, the term “lower alkoxy” applies, as noted above, whereas the term “alkoxy” encompasses “lower alkoxy” as well as alkoxy groups where n is greater than 6 (for example, n=7 to 10). The term “aryloxy” as used herein alone or in combination with another radical means —O-aryl, wherein aryl is defined as noted above.

A protecting group or protective group is a substituent introduced into a molecule to obtain chemoselectivity in a subsequent chemical reaction. Many protecting groups are known in the art and a skilled person would understand the kinds of protecting groups that would be incorporated and could be used in connection with the methods described herein. In “protecting group based peptide synthesis”, typically solid phase peptide synthesis, the desired peptide is prepared by the step-wise addition of amino acid moieties to a building peptide chain. The two most widely used protocols, in solid-phase synthesis, employ tert-butyloxycarbonyl (Boc) or 9-fluorenylmethoxycarbonyl (Fmoc) as amino protecting groups. Amino protecting groups generally protect an amino group against undesirable reactions during synthetic procedures and which can later be removed to reveal the amine. Commonly used amino protecting groups are disclosed in Greene, T. W. et al., Protective Groups in Organic Synthesis, 3rd Edition, John Wiley & Sons (1999). Amino protecting groups include acyl groups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, o-nitrophenoxyacetyl, .alpha.-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, and the like; sulfonyl groups such as benzenesulfonyl, p-toluenesulfonyl and the like; alkoxy- or aryloxy-carbonyl groups (which form urethanes with the protected amine) such as benzyloxycarbonyl (Cbz), p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1-(p-biphenylyl)-1-methylethoxycarbonyl, .alpha.-,.alpha.-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl, t-butyloxycarbonyl (Boc), diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl (Alloc), 2,2,2-trichloroethoxycarbonyl, 2-trimethylsilylethyloxycarbonyl (Teoc), phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl (Fmoc), cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl and the like; aralkyl groups such as benzyl, triphenylmethyl, benzyloxymethyl and the like; and silyl groups such as trimethylsilyl and the like. Amine protecting groups also include cyclic amino protecting groups such as phthaloyl and dithiosuccinimidyl, which incorporate the amino nitrogen into a heterocycle. Typically, amino protecting groups include formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, phenylsulfonyl, Alloc, Teoc, benzyl, Fmoc, Boc and Cbz. It is well within the skill of the ordinary artisan to select and use the appropriate amino protecting group for the synthetic task at hand.

In some embodiments, R¹ is H.

In some embodiments, R² or R³ is covalently linked to R¹ to form proline having NR¹ as the N-terminus.

In some embodiments, R² and R³ are not both H.

In some embodiments, R² and R³ are each independently selected from the group consisting of amino acid chains of a proteinogenic or a non-proteinogenic alpha-amino acids.

In some embodiments, R² and R³ are H and CH₃ respectively or vice versa.

In some embodiments, R² or R³ is —CH2-S—R^(s), wherein R^(s) is selected from lower alkyl; lower amino alkyl; aryl; heteroaryl; alkenyl; or heterocycle; all of which are optionally substituted at one or more substitutable positions with one or more suitable substituents; preferably R^(s) is phenyl or phenyl substituted with lower alkyl, halogen; or lower amino alkyl.

In some embodiments, R⁴ and R⁵ are not both H.

In some embodiments, R** and R*** are not both H.

In some embodiments, R⁴ and R⁵ are each independently H, or C(O)—NHR^(t), wherein R^(t) is H or a lower alkyl. Preferably, R^(t) is tert-butyl or H.

In some embodiments, R⁶ is H.

In some embodiments, R⁶ and either R⁸ or R⁹ form a ring resulting in a proline residue having N—R⁶ as its N-terminus.

In some embodiments, n is 1.

In some embodiments, Z along with L and —C═O is any one of SEQ ID NOs. 1-380.

In some embodiments, X¹ is Leu.

In some embodiments, X² is Asp.

In some embodiments, X³ is Thr.

In some embodiments, X³ is Val.

In some embodiments, X³ is lie.

In some embodiments, X^(y) and X^(z) are each independently a proteinogenic or non-proteinogenic alpha-amino acid.

In some embodiments, X^(z) is a proteinogenic or non-proteinogenic beta-amino acid.

In some embodiments, X^(z) is betaHomoLys or MethylbetaHomoLys.

In some embodiments, X^(y) and X^(z) are each a primary amino acid.

In some embodiments, X^(y) and X^(z) are each any amino acid listed under column X^(y) and column X^(z) respectively of Table 1B.

In various embodiments, the compound is any one of compounds 1-389 and 456 or the multimer is any one of compounds 390-397 and 457-538.

In various embodiments, the multimer is a dimer, trimer, tetramer, or pentamer.

In some embodiments, the monomer compounds are linked by a linker.

In some embodiments, the compounds are linked together at a carbon, nitrogen, oxygen, sulphur or other atom associated with R², R³, R⁴, R⁵, R⁶, R⁷/R⁸, X^(z), or X^(y).

As person skilled in the art would understand that various linkers may be used to multimerize the macrocycles described herein, including esters, amides, amines or mixed amides/amines. Additional linkages include, but are not limited to, ethers, thioethers, thioesters, disulphides, sulfoxides, sulfones, sulfonamides, sulfamates, sulfamides, carbamates, ureas, carbonates, phosphodiesters, phosphonamides, phosphoramidates, heterocycles such as triazoles from azide-alkyne cycloaddition (“Click” chemistry). Alternatively, monomeric macrocycles can be covalently attached to linkers via carbon-carbon single bond linkages, carbon-carbon double bond linkages or carbon-carbon triple bond linkages. Alternatively, monomeric macrocycles can be covalently attached directly to a second, third or fourth monomeric macrocycle via any of the above linkages; in this case no formal linker moiety is present.

In some embodiments, the multimer is a homo-multimer.

In some embodiments, the multimer is a hetero-multimer.

In certain embodiments, there is provided pharmaceutically acceptable salts of the compounds described herein. The term “pharmaceutically acceptable salt,” as used herein, represents salts or zwitterionic forms of the compounds of the present invention which are water or oil-soluble or dispersible, which are suitable for treatment of diseases without undue toxicity, irritation, and allergic response; which are commensurate with a reasonable benefit/risk ratio, and which are effective for their intended use. The salts can be prepared during the final isolation and purification of the compounds or separately by treatment of an amino group with a suitable acid. Representative acid addition salts include acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, formate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethansulfonate (isethionate), lactate, maleate, mesitylenesulfonate, methanesulfonate, naphthylenesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, pectinate, persulfate, 3-phenylproprionate, picrate, pivalate, propionate, succinate, tartrate, trichloroacetate, trifluoroacetate, phosphate, glutamate, bicarbonate, para-toluenesulfonate, and undecanoate. Also, amino groups in the compounds of the present invention can be quaternized with methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dimethyl, diethyl, dibutyl, and diamyl sulfates; decyl, lauryl, myristyl, and steryl chlorides, bromides, and iodides; and benzyl and phenethyl bromides. Examples of acids which can be employed to form therapeutically acceptable addition salts include inorganic acids such as hydrochloric, hydrobromic, sulfuric, and phosphoric, and organic acids such as oxalic, maleic, succinic, and citric. In certain embodiments, any of the peptide compounds described herein are salt forms, e.g., acetate salts.

In an aspect, there is provided, a pharmaceutical composition comprising the multimer described herein along with the pharmaceutically acceptable carrier. The pharmaceutical composition may be formulated for any one of oral delivery, topical delivery and parenteral delivery.

As used herein, “pharmaceutically acceptable carrier” means any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. Examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol and the like, as well as combinations thereof. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Pharmaceutically acceptable carriers may further comprise minor amounts of auxiliary substances such as wetting or emulsifying agents, preservatives or buffers, which enhance the shelf life or effectiveness of the pharmacological agent.

In an aspect, there is provided, a method of treating inflammation or an autoimmune disease in a patient, comprising administering to the patient a therapeutically effective amount of the multimer described herein. Preferably the inflammation or an autoimmune disease is gastrointestinal.

In an aspect, there is provided, a method for treating a condition in a patient associated with a biological function of an α4β7 integrin, the method comprising administering to the patient a therapeutically effective amount of the multimer described herein.

In some embodiments, the condition or disease is Inflammatory Bowel Disease (IBD), ulcerative colitis, Crohn's disease, Celiac disease (nontropical Sprue), enteropathy associated with seronegative arthropathies, microscopic colitis, collagenous colitis, eosinophilic gastroenteritis, radiotherapy, chemotherapy, pouchitis resulting after proctocolectomy and ileoanal anastomosis, gastrointestinal cancer, pancreatitis, insulin-dependent diabetes mellitus, mastitis, cholecystitis, cholangitis, pericholangitis, chronic bronchitis, chronic sinusitis, asthma, primary sclerosing cholangitis, human immunodeficiency virus (HIV) infection in the GI tract, eosinophilic asthma, eosinophilic esophagitis, gastritis, colitis, microscopic colitis, graft versus host disease, colitis associated with radio- or chemo-therapy, colitis associated with disorders of innate immunity as in leukocyte adhesion deficiency-1, chronic granulomatous disease, glycogen storage disease type 1b, Hermansky-Pudlak syndrome, Chediak-Higashi syndrome, and Wiskott-Aldrich Syndrome, or pouchitis resulting after proctocolectomy and ileoanal anastomosis and various forms of gastrointestinal cancer, osteoporosis, arthritis, multiple sclerosis, chronic pain, weight gain, and depression. In another embodiment, the condition is pancreatitis, insulin-dependent diabetes mellitus, mastitis, cholecystitis, cholangitis, pericholangitis, chronic bronchitis, chronic sinusitis, asthma or graft versus host disease.

In preferable embodiments, is an inflammatory bowel disease, such as ulcerative colitis or Crohn's disease.

In an aspect, there is provided, a method for treating a disease or condition in a patient comprising administering to the patient a therapeutically effective amount of the multimer described herein, wherein the disease or condition is a local or systemic infection of a virus or retrovirus.

In some embodiments, the a virus or retrovirus is echovirus 1 and 8, echovirus 9/Barty Strain, human papilloma viruses, hantaviruses, rotaviruses, adenoviruses, foot and mouth disease virus, coxsackievirus A9, human parechovirus 1 or human immunodeficiency virus type 1.

In an aspect, there is provided, a method for treating a disease or condition in a patient comprising administering to the patient a therapeutically effective amount of the multimer described herein, wherein the hepatitis A, B or C, hepatic encephalopathy, non-alcoholic steatohepatitis, cirrhosis, variceal bleeding, hemochromatosis, Wilson disease, tyrosinemia, alpha-1-antitrypsin deficiency, glycogen storage disease, hepatocellular carcinoma, liver cancer, primary biliary cholangitis, primary sclerosing cholangitis, primary biliary sclerosis, biliary tract disease, autoimmune hepatitis, or graft-versus-host disease.

In some embodiments, the multimer inhibits binding of α4β7 integrin to MAdCAM. Preferably, the compound selectively inhibits binding of α4β7 integrin to MAdCAM.

In any embodiment, the patient is preferably a human.

As used herein, the terms “disease”, “disorder”, and “condition” may be used interchangeably.

As used herein, “inhibition,” “treatment,” “treating,” and “ameliorating” are used interchangeably and refer to, e.g., stasis of symptoms, prolongation of survival, partial or full amelioration of symptoms, and partial or full eradication of a condition, disease or disorder in a subject, e.g., a mammal.

As used herein, “prevent” or “prevention” includes (i) preventing or inhibiting the disease, injury, or condition from occurring in a subject, e.g., a mammal, in particular, when such subject is predisposed to the condition but has not yet been diagnosed as having it; or (ii) reducing the likelihood that the disease, injury, or condition will occur in the subject.

As used herein, “therapeutically effective amount” refers to an amount effective, at dosages and for a particular period of time necessary, to achieve the desired therapeutic result. A therapeutically effective amount of the pharmacological agent may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the pharmacological agent to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of the pharmacological agent are outweighed by the therapeutically beneficial effects.

In some embodiments, the compound is administered by a form of administration selected from the group consisting of oral, intravenous, peritoneal, intradermal, subcutaneous, intramuscular, intrathecal, inhalation, vaporization, nebulization, sublingual, buccal, parenteral, rectal, vaginal, and topical.

In some embodiments, the compound is administered as an initial does followed by one or more subsequent doses and the minimum interval between any two doses is a period of less than 1 day, and wherein each of the doses comprises an effective amount of the compound.

In some embodiments, the effective amount of the compound is the amount sufficient to achieve at least one of the following selected from the group consisting of: a) about 50% or greater saturation of MAdCAM binding sites on α4β7 integrin molecules; b) about 50% or greater inhibition of α4β7 integrin expression on the cell surface; and c) about 50% or greater saturation of MAdCAM binding sites on α4β7 molecules and about 50% or greater inhibition of α4β7 integrin expression on the cell surface, wherein i) the saturation is maintained for a period consistent with a dosing frequency of no more than twice daily; ii) the inhibition is maintained for a period consistent with a dosing frequency of no more than twice daily; or iii) the saturation and the inhibition are each maintained for a period consistent with a dosing frequency of no more than twice daily.

In some embodiments, the compound is administered at an interval selected from the group consisting of around the clock, hourly, every four hours, once daily, twice daily, three times daily, four times daily, every other day, weekly, bi-weekly, and monthly.

The compounds described herein may be multimerized using methods and linkers that would be known to a person of skill in the art, for example, as described in WO2016/054411.

The advantages of the present invention are further illustrated by the following examples. The examples and their particular details set forth herein are presented for illustration only and should not be construed as a limitation on the claims of the present invention.

EXAMPLES

Methods and Materials

Synthesis

Methods applicable for making the cyclic peptides described herein can be found generally in Applicant's PCT Publication No. WO 2010/105363 and in U.S.patent application Ser. No. 15/775,319 claiming priority to U.S. Provisional Application No. 62/254,003 filed on Nov. 11, 2015.

More specifically, the below protocols were used to synthesize each of the compounds as indicated in Table 1X. Multimer of the compounds were also synthesized as indicated in Table 2X.

Protocol A: General Nacellin Synthesis

1. Preparation of resin: Fmoc amino acid (1.1 eq. with respect to resin) was dissolved in CH₂Cl₂ (10 mL/g of resin). If the amino acid did not dissolve completely, DMF was added slowly dropwise until a homogeneous mixture persisted upon stirring/sonication. The 2-chlorotrityl resin was allowed to swell in CH₂Cl₂ (5 mL/g of resin) for 15 minutes. The CH₂Cl₂ was then drained and the Fmoc amino acid solution was added to the vessel containing the 2-Cl Trt resin. DIPEA was added (2 eq. with respect to the amino acid) and the vessel was agitated for five minutes. Another 2 eq. of DIPEA was then added and the vessel was left to agitate for an additional 60 minutes. The resin was then treated with methanol (1 mL/g of resin) to endcap any remaining reactive 2-Cl Trt groups. The solution was mixed for 15 minutes, drained and then rinsed with CH₂Cl₂ (×3), DMF (×3), CH₂Cl₂ (×2), and MeOH (×3). The resin was then dried under vacuum and weighed to determine the estimated loading of Fmoc amino acid.

2. Preparation of linear peptide sequence via manual or automated synthesis: Fully protected resin-bound peptides were synthesized via standard Fmoc solid-phase peptide chemistry manually or using an automated peptide synthesizer. All N-Fmoc amino acids were employed.

a. Fmoc deprotection: the resin was treated with 20% piperidine in NMP or DMF twice, for 5 and 10 minutes respectively, with consecutive DMF and NMP washes after each addition.

b. Fmoc amino acid coupling: the resin was treated with 3 eq. of Fmoc amino acid, 3 eq. of HATU and 6 eq. of DIPEA in NMP for 60 minutes. For difficult couplings, a second treatment with 3 eq. of Fmoc amino acid, 3 eq. of HATU and 6 eq. of DIPEA in NMP for 40 minutes was employed.

3. General cleavage with retention of protecting groups: Once the desired linear sequence was synthesized, the resin was treated with either 1.) 1:3, HFIP:CH₂Cl₂ or 2.) 5% TFA in CH₂Cl₂, twice for 30 minutes each, to afford cleavage from the solid support. The solvent was then removed, followed by trituration twice with chilled tert-butyl methyl ether (or diethyl ether/hexanes) to give the desired product. The purity was then analyzed by reverse-phase LCMS.

Protocol B: Preparation of N-Alkylated Fmoc Amino Acid Building Blocks

1. Resin prep: see protocol A, step 1

2. Fmoc deprotection: see protocol A, step 2a

3. Nosyl protection: The deprotected resin was stirred in CH₂Cl₂ (5 mL/mmol of resin) and DIPEA (6.5 eq.). A solution of Nosyl chloride (4.0 eq.) was added slowly, dropwise, over 30 minutes, to avoid a rapid exothermic reaction. After the addition was complete, stirring was continued at room temperature for three hours. The resulting nosyl-protected resin was filtered and washed with CH₂Cl₂, MeOH, CH₂Cl₂, and THF.

4. N-Methylation: To a suspension of resin in THF (10 mL/mmol of resin) was added a solution of triphenylphosphine (5 eq.) in THF (2 M) and MeOH (10 eq.). The stirring suspension was cooled in an ice bath. A solution of DIAD (5 eq.) in THF (1 M) was added dropwise, via addition funnel. After addition was complete the bath was removed and the reaction was stirred at room temperature for an additional 90 minutes. The resin was filtered, washed with THF (×4), CH₂Cl₂ (×3), and THF (×2).

5. Nosyl-deprotection: To a suspension of resin in NMP (10 mL/mmol of resin) was added 2-mercaptoethanol (10.1 eq.) and DBU (5.0 eq.). The solution became a dark green colour. After five minutes, the resin was filtered, washed with DMF until washes ran colourless. This procedure was repeated a second time, and the resin was then washed a final time with CH₂Cl₂.

6. Fmoc protection: To a suspension of resin in CH₂Cl₂ (7 mL/mmol of resin) was added a solution of Fmoc-Cl (4 eq.) in CH₂Cl₂ (7 mL), and DIPEA (6.1 eq.). The suspension was stirred at room temperature for four hours then filtered and washed with CH₂Cl₂ (×2), MeOH (×2), CH₂Cl₂ (×2), and Et₂O (×2).

7. Cleavage from resin: see protocol A, step 3

Protocol C: Reductive Amination

1. Fmoc Weinreb amide formation: a mixture of Fmoc amino acid (1 mmol), N,O-dimethylhydroxylamine.HCl (1.2 eq.), and HCTU (1.2 eq.) in CH₂Cl₂ (6.5 mL), was cooled to 0° C. DIPEA (3 eq.) was then slowly added to the stirring mixture. The cooling bath was removed and the reaction was stirred at room temperature for 16 h. A 10% solution of HCl (4 mL) was added resulting in the formation of a precipitate, which was removed through filtration. The filtrate was washed with 10% HCl (3×4 mL) and brine (2×4 mL). The organic phase was then dried over Na₂SO₄. The solvent was removed under reduced pressure to give crude Fmoc Weinreb amide, which was used in the next reaction without purification.

2. a) Fmoc amino aldehyde formation: lithium aluminum hydride powder (1.5 eq) was placed in a dry flask. THF (Sigma-Aldrich, 250 ppm of BHT, ACS reagent >99.0%, 6.5 mL) was added, and the resulting slurry was cooled to −78° C., with stirring. To the slurry was added a solution of the Fmoc Weinreb amide in THF (10 mL). The reaction vessel was transferred to an ice/water bath, and maintained at 0° C. for 1 h. To the reaction at 0° C., was added dropwise acetone (1.5 mL), then H₂O (0.25 mL) and then the reaction was left to stir for an additional hour at room temperature. The mixture was filtered through Celite, washed with EtOAc (10 mL) and MeOH (10 mL), and the filtrate was concentrated. The crude material was dissolved in CHCl₃ (6.5 mL) and washed with brine (2×3 mL) and the organic phase was then dried over Na₂SO₄, filtered and concentrated to give the Fmoc amino aldehyde.

Alternatively, b) Under argon atmosphere a Lithium Aluminum Hydride 1.0 M solution in THF (Sigma-Aldrich, 157.81 mL, 157.82 mmol, 1 eq.) was slowly added over a solution of the Weinreb amide (157.82 mmol) in THF (Sigma-Aldrich, 250 ppm of BHT, ACS reagent >99.0%, 1 L) at 0° C. and then stirred for 1 h. The reaction at 0° C., was diluted with Et₂O (500 mL) and the resultant solution was washed with 10% NaHSO₄ (10×300 mL), 10% KHSO₄ (10×300 mL) and HCl (10×300 mL). The organic phase was then dried over Na₂SO₄, filtered and concentrated to afford the crude Fmoc amino aldehyde.

3. Reductive amination on-resin: the linear peptide on-resin was placed in a solid-phase peptide synthesis reaction vessel and diluted with DMF (22 mL/g of resin). The Fmoc aldehyde (4.0 eq.) was added and the reaction was left to shake overnight. The solution was then drained and the resin was washed with CH₂Cl₂ (×3) and DMF (×3). The resin was then diluted with a mixture of MeOH/CH₂Cl₂ (22 mL/g of resin, 1:3 ratio) and NaBH₄ (7 eq.) was subsequently added. The mixture was left to shake for four hours, then the solution was drained and the resin was washed with CH₂Cl₂ (×3) and DMF (×3).

Protocol D: Fragment-Based Macrocyclization

a) In a two-dram vial, 0.1 mmol of the linear peptide and DEPBT (1.5 eq.) were dissolved in 5 mL of freshly distilled THF (0.02 M). DIPEA (3 eq.) was then added and the reaction mixture was left to stir overnight at room temperature (16 h). Tetraalkylammonium carbonate resin (Biotage®, 6 eq.) was then added to the reaction mixture and stirring was continued for an additional 24 h. The reaction was then filtered through a solid-phase extraction vessel and rinsed with CH₂Cl₂ (2 mL). The filtrate and washes were combined and the solvent was removed under reduced pressure.

Alternatively, b) In a two-dram vial, 0.1 mmol of the linear peptide and HATU (2.0 eq.) were dissolved in 80 mL of CH₂Cl₂ (1.25 mM). DIPEA (6 eq.) was then added and the reaction mixture was left to stir overnight at room temperature (16 h). The solvent was removed under reduced pressure.

Protocol E: Aziridine Aldehyde-Based Macrocyclization

The linear peptide was dissolved in TFE (if solubility problems were encountered, a 50:50 mixture of TFE:CH₂Cl₂ was used for the cyclization). Then 0.6 eq. of (S)-aziridine-2-carboxaldehyde dimer (prepared as per literature protocol: J. Am. Chem. Soc. 2006, 128 (46), 14772-14773 and Nat. Protoc. 2010, 5 (11), 1813-1822) as a TFE stock solution (0.2 M) was added, giving a final reaction mixture concentration of 0.1 M. tert-Butyl isocyanide (1.2 eq.) was then added and the reaction mixture was stirred for four hours. Progress was analyzed along the way via LC-MS.

Protocol F: Nucleophilic Ring-Opening of Acyl Aziridine, Post Macrocyclization

a) Thioacetic acid/thiobenzoic acid: the corresponding thio acid (4 eq.) was added to the crude reaction mixture. Reaction progress was monitored by LC-MS, and was generally complete after 1-2 hours.

Alternatively, b) Thiophenol: Thiophenol (4 eq.) and DIPEA (4 eq.) were added to the crude cyclization mixture. Reaction progress was monitored by LC-MS, and was generally complete after 1-2 hours. Solvent was removed under reduced pressure and dried under vacuum. Crude material was either triturated with Et₂O/hexanes or TBME, or alternatively, diluted with H₂O, frozen and lyophilized.

Protocol G: Suzuki Coupling, Post Macrocyclization

a) As a general example, an iodo-Phe-containing macrocycle (0.1 mmol), Na₂CO₃ (2 eq.), substituted boronic acid (1.1 eq.) and 4 mL of water:acetonitrile (1:1 ratio) were combined in a microwave vial. The mixture was treated with N₂ gas flow for 10 minutes. While under N₂, silicon based Pd-catalyst (Siliacat-DPP Pd heterogenous catalyst, 0.05 eq.) was added. The reaction vial was sealed and placed in the microwave for 10 minutes at 120° C. (reaction time and temperature were increased to 30 min. and 150° C., depending on the substrate) or thermally heated at 90° C. for 1 h. Reaction progress was monitored by LCMS. Once complete, the reaction was filtered through a Celite plug and the solvent was removed under reduced pressure.

Alternatively, b) as a specific example, Suzuki couplings with macrocycles that were prepared using 3-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-bromobenzoic acid were conducted as follows: A mixture of crude macrocyclic Compound 340 that had been orthogonally protected as the β-tert-butyl ester of the Asp residue and the tert-butyl ether of the Thr residue (200 mg, 0.22 mmol) and 4-(4-Boc-piperazino) phenylboronic acid pinacol ester (171 mg, 0.44 mmol) were dissolved in a 1,2-dimethoxyethane (5.4 mL) and Ethanol (1.2 mL) at room temperature. Water (1.2 mL) was added to the solution, followed by Na₂CO₃ (35 mg, 0.33 mmol). The reaction flask was flushed for at least 5 to 10 min under nitrogen gas and then catalyst SiliaCat-DPP Pd (88 mg, 10 mol %, 0.25 mmol/gm) was added. The reaction mixture was heated with stirring under nitrogen at 90° C. for 1 hr. LCMS after 1 hour showed complete consumption of substrate and ˜5% de-bromination compound; the desired Suzuki cross-coupled product represented ˜84% yield after taking into account the excess of boronate ester by UV. The reaction mixture was cooled to room temperature and filtered over a celite pad to remove catalyst SiliaCat-DPP Pd. The celite pad was washed with a little DCM and the solvents were removed under vacuum to give pale yellow crude solid as the Suzuki coupling product. Reagent 3-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-bromobenzoic acid was itself prepared from methyl 3-(aminomethyl)-4-bromobenzoate (US2011251247) via saponification of the methyl ester and protection of the amine as the Fmoc carbamate, as follows: to a solution of methyl 3-(aminomethyl)-4-bromobenzoate (1.36 g, 5.57 mmol) in Dioxane (33 ml) and Water (9 ml) was added lithium hydroxide (6.13 ml, 6.13 mmol). The mixture was stirred for 3 hrs at room temperature. TLC showed the hydrolysis reaction was complete. Dioxane (16 ml) was added. The mixture was neutralized by the addition of 1 N HCl (aq) (6.17 mL). Sodium bicarbonate (0.468 g, 5.57 mmol) was added, followed by (9H-fluoren-9-yl)methyl carbonochloridate (2.162 g, 8.36 mmol). The mixture was stirred for 2 hrs at room temperature and was acidified to pH 3 by the addition of 1 N HCl (aq) (6.2 mL). Water (40 ml) was added, extracted with AcOEt (4×150 mL). The combined organic layers were dried over sodium sulfate and the solvent was evaporated to 50 ml. Precipitation began to occur and was allowed to slowly continue overnight at room temperature. White solid was then collected by filtration, washed with hexane and dried under high vacuum to afford 3-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-bromobenzoic acid (2.0 g, 4.42 mmol, 79% yield).

Protocol H: General Ulmann Coupling, Post Macrocyclization

Under inert atmosphere, the peptide macrocycle (0.018 mmol) was placed in a 2-dram vial containing 2 mL of dry CH₂Cl₂. Cu(OAc)₂ (1 eq.), benzene boronic acid (2 eq.) and 4 Å (oven-dried) molecular sieves were then added to the vial followed by DIPEA (4 eq.). The contents of the vial were stirred at room temperature overnight. The reaction progress was assessed by LCMS. Once the reaction was deemed complete, the mixture was filtered through a Celite plug and the solvent was removed under reduced pressure.

Protocol I: General Global Deprotection and Cleavage

Deprotection of the side chain protecting groups was achieved by dissolving the peptides in 2 mL of a cleavage cocktail consisting of TFA:H₂O:TIS (95:2.5:2.5) for two hours (for sensitive peptides the mixture of TFA:H₂O:TIS (95:2.5:2.5) may be substituted for a mixture of TFA:CH₂Cl₂ (50:50)). Subsequently, the cleavage mixture was evaporated under reduced pressure and the peptides were precipitated twice from chilled diethyl ether/hexanes (or tert-butyl methyl ether).

Protocol J: General Cleavage of Reductively-Labile Protecting Groups

a) Pd/C and formic acid debenzylation: the benzyl protected macrocycle (0.35 mmol) was dissolved in MeOH (8 mL) with 10% formic acid, 10% wt. Pd/C (Sigma-Aldrich, 37 mg, 0.1 Eq) and heated to 55° C. for 1 h to 4 h. Once the reaction was deemed complete, the mixture was filtered through a Celite plug, washed with MeOH and the solvent was removed under reduced pressure.

Or alternatively, b) Raney Ni desulfurization/debenzylation: Raney Ni slurry (1-2 mL) was added directly to the cyclization reaction mixture and stirred vigorously overnight. The vial was then centrifuged and the liquid was transferred using a pipette to a tared vial. MeOH was added to the vial containing Raney Ni. The vial was then sonicated, vortexed, and centrifuged. Again, the liquid was transferred to a tared vial. This process was repeated with EtOAc and then a final time with MeOH. The combined washes were then removed under reduced pressure and the residue dried under vacuum.

Protocol K: Amidation of Side Chain, Post Macrocyclization

Macrocycle (0.021 mmol) was dissolved in 1 mL of CH₃CN. K₂CO₃ (5 eq.) and the corresponding acid chloride (2 eq.) were then added and the reaction mixture was left to stir at room temperature overnight. Reaction progress was checked by LC-MS in the morning. Upon completion, the solvent was removed by reduced pressure.

Protocol L: Fluorescent Dye Attachment

The macrocycle (4 μmol) was dissolved in DMSO (200 μL). DIPEA (5 eq.) was then added. In a separate vial, 5 mg of fluorescent dye as the NHS ester was dissolved in 200 μL of DMSO. The macrocycle solution was then added to the solution of the fluorescent label. The reaction mixture was stirred overnight. Reaction progress was checked by LC-MS in the morning and then the solvent was removed by lyophilization.

Protocol M: Purification Methods

All macrocycles were purified using reverse-phase flash column chromatography using a 30 g RediSep C18 Gold Column. The gradient consisted of eluents A (0.1% formic acid in double distilled water) and B (0.1% formic acid in HPLC-grade acetonitrile) at a flow rate of 35 mL/min.

Multimerization Protocols

Protocol N: Linker Synthesis for Multimerization

a) Preparation of Acyl chloride linkers: Di-, tri- or tetra-carboxylic acids (1 eq.) and CH₂Cl₂ (0.114 M concentration) were added to a two-dram vial. SOCl₂ (15 eq. per carboxylic acid) was then added and the reaction mixture was left to stir for four hours at room temperature (some substrates required heating at 70° C. overnight for full solution and/or conversion). The solvent was removed via N₂ flow. The residue was dissolved in 3 mL of dry CH₂Cl₂ which was then removed under N₂ flow. This process was performed two additional times in an attempt to remove any free HCl from the sample. The resulting residue was then used without purification in the dimerization reaction.

b) Preparation of Benzotriazole linkers, Method A: Thionyl chloride (2 eq. per carboxylic acid) was added to a solution of benzotriazole (10 eq. per carboxylic acid) in dichloromethane (20 mL per mmol of starting linker) and the solution was stirred at room temperature for 20 min. The di-, tri- or tetra-carboxylic acids (1 eq.) were added to each mixture, which were then stirred at room temperature for 24 h (a change in order of addition did not materially alter the outcome). The reaction was quenched with NaHCO₃ (10%, 100 mL) and the layers were separated. The organic layer was washed with HCl (10%, 2×100 mL) and NaHCO₃ (10%, 2×100 mL), dried over anhydrous sodium sulfate, filtered and evaporated under vacuum to give the desired Benzotriazole-activated carboxylic acids.

c) Preparation of Benzotriazole linkers, Method B: To a suspension of HATU (1.5 eq. per carboxylic acid), Benzotriazole (2 eq. per carboxylic acid) and the di-, tri- or tetra-carboxylic acids (1 eq.) in dichloromethane (20 mL per mmol of starting linker) was added DIPEA (3 eq. per carboxylic acid) and the resultant yellow solution was stirred at room temperature for 16 h. The reaction was quenched with NaHCO₃ (10%, 100 mL) and the layers were separated. The organic layer was washed with HCl (10%, 2×100 mL) and NaHCO₃ (10%, 2×100 mL), dried over anhydrous sodium sulfate, filtered and evaporated under vacuum to give the desired Benzotriazole-activated carboxylic acids

d) Preparation of Lys(CBz)-Pimelic acid-Lys(CBz) linker: Pimelic acid was converted to the bis-Benzotriazole-activated moiety using Protocol Nb. Commercial/VP-Z-L-lysine methyl ester hydrochloride (2 eq.; Chemlmpex) was treated with bis-Benzotriazole-activated Pimelic acid (1 eq.) in CH₃CN (0.011 M) containing DIPEA (10 eq.). The reaction mixture was stirred for 16 h (monitored by LC-MS). The solvent was removed by rotoevaporation and the crude material was submitted to reverse-phase silica chromatography (Biotage) to obtain the purified bis-methyl ester of Lys(CBz)-Pimelic acid-Lys(CBz) as an intermediate. To a solution of the bis-methyl ester (1.5 mmol, 1.0 eq.) in THF (10 mL) were added LiCl (3.0 mmol, 2.0 eq.) and LiOH—H₂O (3.0 mmol, 2.0 eq.), followed by H₂O (250 uL) to help solubilize the salts. The reaction was stirred at room temperature overnight. Upon completion of the hydrolysis, as assessed by LC-MS monitor, formic acid was added dropwise to neutralize the basic solution. The solvent was removed by rotoevaporation and the crude material was submitted to reverse-phase chromatography (Biotage) to obtain the purified di-acid linker.

e) Preparation of PEG2-Diglycolic acid-PEG2 linker: Diglycolyl chloride (0.35 mmol; 1 eq.; Sigma Aldrich cat. No. 378151) in anhydrous CH₂Cl₂ (5 mL) was treated with NH₂-PEG2-CH₂CH₂COOtBu (2 eq.; Biochempeg Cat. No. MD005067-2), followed by dropwise addition of DIPEA (3.5 mmol, 10.0 eq); NB— this order of addition proved to be very important. The reaction was monitored by LC-MS. After 30 min., the reaction was complete, and longer stirring times did not affect the product ratio. The solvents were removed in vacuo and the crude material was submitted to reverse-phase chromatography (Biotage) to obtain the purified di-tert-butyl ester intermediate. Removal of the tert-butyl ester groups was effected by Protocol I. The diacid linker was isolated as a crude and used as such multimerization reactions without further manipulation.

f) Preparation of PEG2-Diphenic acid-PEG2 linker: Diphenic acid was converted to the bis-Benzotriazole-activated moiety using Protocol Nb. Commercial NH₂-PEG2-CH₂CH₂COOtBu (2 eq.; Biochempeg Cat. No. MD005067-2) was treated with bis-Benzotriazole-activated Diphenic acid (1 eq.) in CH₃CN (0.011 M) containing DIPEA (10 eq.). The reaction mixture was stirred for 16 h (monitored by LC-MS). The solvent was removed by rotoevaporation and the crude material was submitted to reverse-phase silica chromatography (Biotage) to obtain the purified di-tert-butyl ester intermediate. Removal of the tert-butyl ester groups was effected by Protocol I. The diacid linker was isolated as a crude and used as such in multimerization reactions without further manipulation.

g) Preparation of PEG2-Pimelic acid-PEG2 linker: Pimelic acid was converted to the bis-Benzotriazole-activated moiety using Protocol Nb. Commercial NH₂-PEG2-CH₂CH₂COOtBu (2 eq.; Biochempeg Cat. No. MD005067-2) was treated with bis-Benzotriazole-activated Pimelic acid (1 eq.) in CH₃CN (0.011 M) containing DIPEA (10 eq.). The reaction mixture was stirred for 16 h (monitored by LC-MS). The solvent was removed by rotoevaporation and the crude material was submitted to reverse-phase silica chromatography (Biotage) to obtain the purified di-tert-butyl ester intermediate. Removal of the tert-butyl ester groups was effected by Protocol I. The diacid linker was isolated as a crude and used as such in multimerization reactions without further manipulation.

Protocol O: Nacellin Multimerization

a) Multimerization of amine-containing monomeric macrocycles using bis- or tris-acyl chloride-activated linkers: The corresponding acyl chloride (0.35 mmol, 1.0 eq.), freshly prepared and under Argon atmosphere, was dissolved in anhydrous CH₂Cl₂ (5 mL; note that larger scale reactions required more-concentrated solution to produce higher-yielding dimerizations). Monomeric macrocycle (2, 3 or 4 eq. for bis-, tris-, or tetra-acyl chlorides), optimally supplied as the free-base/non-salted form of the reacting amine center, was added to the flask, followed by dropwise addition of DIPEA (3.5 mmol, 10.0 eq); NB—this order of addition proved to be very important. The reaction was monitored by LC-MS. After 30 min., the reaction was complete, and longer stirring times did not affect the product ratio. The solvents were removed in vacuo and the crude material was submitted to reverse-phase chromatography (Biotage) to obtain the purified product.

b) Multimerization of amine-containing monomeric macrocycles using Benzotriazole-activated linkers: To a solution of monomeric macrocycle (2, 3 or 4 eq.), optimally supplied as the free-base/non-salted form of the reacting amine center, and the corresponding Benzotriazole-activated linker, previously prepared but not longer than 1 week prior to multimerization, (0.011 mmol, 1 eq.) in CH₃CN (1 mL) in the presence of DIPEA (0.02 mL, 0.114 mmol, 10 eq). The reaction mixture was stirred for 16 h (monitored by LC-MS). The solvent was removed by rotoevaporation and the crude material was submitted to reverse-phase silica chromatography (Biotage) to obtain the purified product.

c) Dimerization of amine-containing monomeric macrocycles using 2-Chloroacetyl chloride: To a solution of the monomeric macrocycle (0.0571 mmol, 2 eq.), optimally supplied as the free-base/non-salted form of the reacting amine center, in distilled THF (1.0 mL), were added 2-chloroacetyl chloride (3.19 mg, 0.029 mmol, 1 eq.) followed by DIPEA (25 uL, 0.17 mmol, 6.0 eq.). The reaction mixture was stirred for 16 h (monitored by LC-MS). NaI (8.5 mg, 0.05708 mmol, 2 eq) was then added and the reaction mixture was heated at 50° C. for 2 h. The solvent was removed in vacuo and the crude material was submitted to reverse-phase silica chromatography (Biotage) to obtain the purified product.

d) Dimerization of amine-containing monomeric macrocycles using Acryloyl chloride: To a solution of the monomeric macrocycle (0.0571 mmol, 2 eq.), optimally supplied as the free-base/non-salted form of the reacting amine center, in distilled THF (1.0 mL), were added Acryloyl chloride (2.6 mg, 0.029 mmol, 1 eq.) and then DIPEA (25 uL, 0.17 mmol, 6.0 eq.). The reaction mixture was stirred for 16 h (monitored by LC-MS). DBU (8.5 uL, 0.057 mmol, 2 eq) was then added and the reaction was heated at 50° C. for 5 h. The solvent was removed in vacuo and the crude material was submitted to reverse-phase silica chromatography (Biotage) to obtain the purified product.

e) Multimerization of hydoxyl-containing monomeric macrocycles: Di-, tri- or tetra-carboxylic acid linker (4.3 μmol), monomeric macrocycle (2, 3 or 4 eq.), DMAP (2, 3 or 4 eq.), and EDC.HCl (4, 8 or 12 eq.) were dissolved in DCM (500-1000 μL). The reaction mixture was left to stir at room temperature overnight. Reaction progress was assessed by LC-MS. Upon completion, the solvent was removed under reduced pressure and the crude was submitted to reverse-phase silica chromatography (Biotage) to obtain the purified product.

f) Dimerization of amine-containing monomeric macrocycles using 2,4-dichloro-5-nitropyrimidine: To a solution of 2,4-dichloro-5-nitropyrimidine (2.0 mg, 0.010 mmol, 1.0 equiv) and monomeric macrocycle (0.021 mmol, 2.1 eq.), optimally supplied as the free-base/non-salted form of the reacting amine center, in chloroform (1 mL), in a 1-dram vial, was added DIPEA (0.02 mL, 0.11 mmol, 11.0 equiv); the reaction mixture immediately turned yellow. Stirring was continued at room temperature overnight, at which point LC-MS analysis exhibited almost full conversion to desired dimer. An additional 24 h of reaction time did not lead to any further conversion. Solvent was rotoevaporated to dryness, and the crude residue was submitted to reverse-phase chromatography to afford the purified material in 76% isolated yield.

g) Multimerization of amine-containing monomeric macrocycles using HATU-activated linkers: To a solution of the monomeric macrocycle (2, 3 or 4 eq.), optimally supplied as the free-base/non-salted form of the reacting amine center, in 1 mL dry DCM, was added the di-, tri- or tetra-substituted carboxylic acid (1 eq.) under inert atmosphere at room temperature. HATU (3, 6 or 9 eq.) was added to the solution, followed by the addition of DIPEA (3, 6 or 9 eq.). The reaction mixture was left to stir overnight. Assessment of reaction progress by LC-MS after 14 h indicated completion. The reaction mixture was rotoevaporated to near-dryness, then placed under high vacuum. If no orthogonal protecting groups required removal (for example, amines protected as the CBz carbamate), the crude material was submitted to reverse-phase chromatography to afford the purified material.

h) Multimerization of amine-containing monomeric macrocycles using halide-activated linkers: To a solution of monomeric macrocycle (3.0 eq. if used with a dihalide, 4.5 eq. if used with a trihalide) and the corresponding di or tri-halide linker (1.0 eq) in CH₃CN (2 mL) was added DIPEA (˜30 eq.). The reaction mixture was stirred for 16 h (monitored by LC-MS). The solvent was removed, and crude was submitted to reverse-phase chromatography to afford the purified material.

Integrin α4β7-MAdCAM-1 ELISA Competition Assay

A 96-well Microlon plate (Greiner, 655001) was coated with 100 μl per well of a solution of 1 μg/ml recombinant integrin α4β7 (R&D Systems, 5397-A3-050) in carbonate buffer (50 mM, pH 9.6). The plate was incubated at 4° C. overnight. The solution was removed and 250 μl blocking buffer (50 mM Tris, 150 mM NaCl, 1 mM MnCl₂, 1% BSA, 0.05% Tween) was added per well. The plate was then incubated for 1 hour at room temperature. The plate was washed three times with wash buffer (50 mM Tris, 100 mM NaCl, 1 mM MnCl₂, 0.05% Tween). To each well, 50 μl of compound diluted in assay buffer was added by transfer from a compound serial dilution plate. 50 μl recombinant MAdCAM-Fc (R&D systems, 6056-MC-050) at a concentration of 0.1 μg/ml in assay buffer (50 mM Tris, 150 mM NaCl, 1 mM MnCl₂, 0.1% BSA, 0.05% Tween) was added to each well. The plate was incubated at room temperature with shaking (300 rpm) for 2 hours to reach binding equilibrium. Then the plate was washed three times in wash buffer and 100 μl anti-human IgG Fc specific-HRP (Abcam, Ab97225) diluted at 1:2000 in assay buffer was added to each well. The plate was incubated at room temperature for 1 hour under agitation. The plate was then washed three times and 100 μl of 1,3′,5,5′-Tetramethylbenxidie (TMB, KPL 5120-0083) was then added to each well. The reaction was stopped after 2 minute-incubation by adding 50 μl of 1M H₂SO₄ and optical absorbance was read at 450 nM.

Integrin α4β1-VCAM-1 Competition ELISA

A 96-well Microlon plate (Greiner, 655001) was coated with 100 μl per well of a solution of 0.5 g/ml recombinant integrin α4β1 (R&D Systems, 5397-A3-050) in carbonate buffer (50 mM, pH 9.6). The plate was incubated at 40° C. overnight. The solution was removed and 250 μl blocking buffer (50 mM Tris, 150 mM NaCl, 1 mM MnCl₂, 1% BSA, 0.05% Tween) was added per well. The plate was then incubated for 1 hour at room temperature. The plate was washed three times with wash buffer (50 mM Tris, 100 mM NaCl, 1 mM MnCl₂, 0.05% Tween). To each well, 50 μl of compound diluted in assay buffer was added by transfer from a compound serial dilution plate. 50 μl recombinant VCAM-Fc (R&D systems, 862-VC-100) at a concentration of 0.1 μg/ml in assay buffer (50 mM Tris, 150 mM NaCl, 1 mM MnCl₂, 0.1% BSA, 0.05% Tween) was added to each well. The plate was incubated at room temperature with shaking (300 rpm) for 2 hours to reach binding equilibrium. Then the plate was washed three times in wash buffer and 100 μl anti-human IgG Fc specific-HRP (Abcam, Ab97225) diluted at 1:2000 in assay buffer was added to each well. The plate was incubated at room temperature for 1 hour under agitation. The plate was then washed three times and 100 μl of 1,3′,5,5′-Tetramethylbenxidie (TMB, (TMB, KPL 5120-0083) was then added to each well. The reaction was stopped after 2 minute-incubation by adding 50 μl of 1M H₂SO₄ and optical absorbance was read at 450 nM.

Integrin α4β7-MAdCAM Cell Adhesion Assay

RPMI8866 human cells (Sigma #95041316) were cultured in RPMI 1640 medium (HyClone SH30027.1) supplemented with 10% FBS (Seradigm) and 1% Penicillin-Streptomycin. A 96-well plate (Costar, 3603) was coated with 100 ml/well of human recombinant MAdCAM-1 Fc Chimera (R&D Systems, 6056-MC-050) solution at 0.25 μg/ml in coating buffer (50 mM sodium carbonate, pH 9.6). The plate was incubated overnight at 4° C. and washed twice with 150 μl per well wash buffer (0.05% Tween 20 in PBS), blocked with 250 μl per well blocking buffer (1% non-fat dry milk in PBS), and incubated for 2 hours at room temperature. RPM18866 cells were resuspended at 10 million cells/ml in PBS containing 5 mM calcein and incubated at 37° C. for 30 min in a 50 ml tube. PBS was added to fill the tube, cells were spun down and resuspended in RPMI 1640 medium to 2 million/ml. Compounds were diluted by serial dilution in binding buffer (1.5 mM CaCl₂, 0.5 mM MnCl₂, 50 mM Tris-HCl, pH 7.5) to a final volume of 50 μl per well at 2× concentration. The plate was washed once with 300 □l of PBS, 50 μl of compound and 50 μl of cells (100,000 cells) were transferred to each well and the plate was incubated in the dark at 37° C., 5% CO₂ for 45 min to allow cell adhesion. The plate was emptied by inverting and blotting on paper towels and washed manually twice with PBS. 100 μl PBS was then added to each well. The fluorescence was read (Ex₄₉₅/Em₅₁₅) using a plate reader (Tecan Infinite 1000). To calculate the dose response, the fluorescence value of control wells not containing cells was subtracted from each test well.

Integrin α4β1-VCAM Cell Adhesion Assay

RAMOS human cells (ATCC CRL-1596) were cultured in RPMI 1640 medium (HyClone SH30027.1) supplemented with 10% FBS (Seradigm) and 1% Penicillin-Streptomycin. A 96-well plate (Costar, 3603) was coated with 100 ml/well of recombinant human VCAM-1 Fc Chimera (R&D systems, 862-VC-100) solution at 0.25 μg/ml in coating buffer (50 mM sodium carbonate, pH 9.6). The plate was incubated overnight at 4° C. and washed twice with 150 μl per well wash buffer (0.05% Tween 20 in PBS), blocked with 250 □l per well blocking buffer (1% non-fat dry milk in PBS), for 1 hour at room temperature. During blocking step, RAMOS cells were resuspended at 10 million cells/ml in PBS containing 5 mM calcein and incubated at 37° C. for 30 min in a 50 ml tube. PBS was added to fill the tube, cells were spun down and resuspended in RPMI 1640 medium to 2 million/ml. Compounds were diluted by serial dilution in binding buffer (1.5 mM CaCl₂, 0.5 mM MnCl₂, 50 mM Tris-HCl, pH 7.5) to a final volume of 50 μl per well at 2× concentration. The plate was washed once with 300 μl of PBS, 50 μl of compound and 50 μl of cells (100,000 cells) were transferred to each well and the plate was incubated in the dark at 37° C., 5% CO₂ for 45 min to allow cell adhesion. The plate was emptied by inverting and blotting on paper towels and washed manually twice with PBS. After last wash, 100 μL of PBS was added to wells and the fluorescence was read (Ex₄₉₅/Em₅₁₅) using a plate reader (Tecan Infinite 1000). To calculate the dose response, the fluorescence value of control wells not containing cells was subtracted from each test well.

Analyte Competition Assay in CD4+ Integrin α₄+β₇-lo Memory T Cells

Receptor occupancy in primary cells was determined by measuring the amount of biotinylated human recombinant MAdCAM-1-FC or human recombinant VCAM-1-Fc bound to selected cell populations using flow cytometry. Human recombinant MAdCAM-1-FC or human recombinant VCAM-1-FC (R&D systems) were biotinylated using commercially available reagents and protocol (Pierce).

Whole blood was collected from human donors in sodium heparin tubes. A volume of 100 microL of blood was incubated with compound and 4 mM MnCL₂ for 1 hour at room temperature. Cells were washed twice with 1 mL of 1×DPBS calcium magnesium free (CMF) (ThermoFisher Scientific) and resuspended in 100 microL of DPBS CMF.

Biotinylated human recombinant MAdCAM-1-Fc or VCAM-1-Fc were added at saturating concentration and incubated at room temperature for 1 hour. A volume of 2 mL of 1×BD FACS Lyse (BD Biosciences) was then added and the mixture was incubated for 8-12 minutes at room temperature in the dark to lyse red blood cells. Cells were washed with 1 mL stain buffer-FBS (BD Biosciences) and resuspended in 100 μl stain Buffer-FBS (BD Biosciences) containing 4 mM MnCl₂. Biotinylated-rhMAdCAM-1 was applied at a saturating concentration of 1200 ng/mL to compete with test article binding and incubated at room temperature for 1 hour. Cells were then washed with 1 mL stain buffer-FBS and resuspended in 100 μl stain buffer-FBS. The cells were incubated in the dark for 30 minutes at room temperature with 1 ul Streptavidin APC (Biolegend 0.2 mg/ml) and a panel of antibodies for the detection of memory T helper α4β7-positive cells subset. And amount of 5.0 ul each of the following antibodies were used; CD45 FITC (BioLegend 200 ug/ml), CD29 APC Cy7 (BioLegend 100 ug/ml), Integrin beta7 PE, (BioLegend concentration 50 μg/mL), CD49d V421 (BioLegend 50 μg/mL), CD3 V510 (BioLegend 30 μg/mL), CD4 PECy7 (BioLegend 100 μg/mL), CD45RO PerCP, BioLegend 200 μg/mL). The cells were then washed with stain-buffer-FBS and resuspended in 150 microL stain buffer-FBS for acquisition on the flow cytometer (BD FACSCanto™ flow cytometer and BDFACSDiva™ software). FACS data was acquire by electronic gating on the basis of forward versus side scatter, The cytometer was set to collect 20,000 events in each tube. Cell population were determined using the following markers, CD45+, CD3+, CD4+, CD45RO+, CD49d+, integrin b7, biotinylated ligands.

Compound receptor occupancy was defined as the decrease in the number of integrin β₇+ or integrin β₇-lo cells binding biotinylated rhMAdCAM-1 or rhVCAM-1, respectively.

Receptor occupancy was calculated with the following equation: 100−((% ligand-positive cells with compound/% ligand-positive cells DMSO)*100)

In vivo T lymphocyte trafficking analysis in mouse model of colitis

Animal care: The animal care facility employed is accredited by the Canadian Council on Animal Care (CCAC). This study was approved by a certified Animal Care Committee and complied with CACC standards and regulations governing the use of animals for research. The animals were housed under standardized environmental conditions. A standard certified commercial rodent diet was provided ad libitum. Tap water was provided ad libitum at all times.

Dextran sulfate sodium (DSS) was administered to C57Bl/6 female mice for five days through addition to their drinking water at 3%. Body weight and disease activity index (“DAI”) were measured on Day 5 in order to distribute DSS-treated animals in uniform groups prior to dosing. DAI was scored based on the severity three specific symptoms associated with colitis: 1—blood in stool (negative hemoccult, positive hemoccult, blood traces in stool visible, rectal bleeding); 2—stool consistency (normal, soft but still formed, very soft, diarrhea); 3—body weight loss.

From Day 6 to day 9, Compound No. 517 (ET03764) or the vehicle were administered orally daily at 5 ml/kg. On day 9, four hours after dosing, the animals were euthanized by cardiac puncture under general anesthesias. Mesenteric lymph nodes (MLN) were collected, triturated, and washed in HBSS-FCS. The cells were incubated for 15 minutes in BD mouse FcBlock followed by 30-minute incubation with specific antibodies. After washes, cells were either fixed using BD fix solution or immediately process for cell surface marker staining. The antibodies used were as followed: CD4 PE (BD Bioscience), CD44 FITC (BD Biosciences), CD45RB PerCy 5.5 (BD Biosciences), α4β7 PE (eBiosciences). Cell populations were then analyzed using FACSCanto cytometer and gating on CD4+, CD44^(hi), CD45RB^(low), α4β7+.

Statistical analysis was performed using GraphPad Prism. Differences among groups were evaluated by two-way ANOVA, wit a 95% confidence interval.

Results and Discussion

Compounds were synthesized in accordance with the above-noted methods. A selection of compounds was characterized using NMR (not all data shown). A subset of NMR data is provided in FIG. 6 for Compound No. 390.

Binding Affinity and Selectivity of Compounds for Integrin α4β7 and α4β1

We measured binding potency for monomeric and dimeric compounds to α4β7-integrin using a battery of biochemical, cell-based and ex-vivo assays. Multimeric compounds were generally more potent in cellular assays.

We measured the ability of test articles to prevent the adhesion of RMP18866 cells, which express integrin α4β7, to plates coated with MAdCAM-1. Multimeric compounds were generally more potent in their ability to inhibit cell adhesion than their constituent monomers. For example Compound No. 340 (ET2451) and Compound No. 456 (ET4062) had IC50 of 175 and 199 nM respectively in the RPM18866 cell adhesion assays (Table 1C and 1C′). Multimeric compounds with over 10-fold greater potency than their constituent monomeric compounds were generated. For example, Compound No. 517 (ET3764), a homodimer of Compound No. 340 (ET2451), had an IC50 of 9.9 nM in the RPM18866 cell adhesion assay. Compound multimers generated from monomeric Compound 456 (ET4062) also showed higher binding affinity (Table 2C).

Similar results were obtained in a ligand competition assay for binding to integrin α4β7 in human whole blood. Receptor occupancy of nacellins was determined by measuring the proportion of α4β7+ memory T helper cells able to bind biotinylated rhMAdCAM-1 using flow cytometry (FIG. 7). Multimeric compounds were able to compete with MadCAM-1 on α4β7-positive primary cells with greater potency than monomeric compounds. Two general monomeric chemotypes were shown to compete more effectively, with increased potency for binding to integrin α4β7 when multimerized using a variety of linkers. For example, Dimeric Compound No.s 534 (ET4113) and 535 (ET4110) demonstrated IC50 of 38 and 76 nM respectively while the corresponding parent monomeric Compound No. 456 (ET4062) only reached 15% receptor occupancy at the maximum concentration of 1000 nM. Similarly, the dimeric Compound No.s 517 (ET3764) and 390 (ET3755) competed with a saturating amount of MAdCAM, with EC50 of 38 and 90 nM respectively within the same study. The corresponding monomeric Compound 340 (ET2451) reached 50% receptor occupancy at low concentrations but no concentration-response curve could be obtained. This could be the result of non-specific binding of the monomeric compound to the cell.

Interestingly, differences in binding affinity between monomeric and multimeric compounds were not as pronounced in ELISA binding assays. It is possible that avidity enhances the binding potency of multimeric compounds in cells.

Multimeric compounds showed enhanced selectivity for integrin α4β7 over integrin α4β1. In order to determine the selectivity of the compounds in cell assays, we measured the adhesion of Ramos cells, which express integrin α4β1 to VCAM-coated plates. Multimeric compounds had generally higher selectivity for integrin α4β7 over integrin α4β1 than their monomeric constituents. For example, monomeric Compound No.s 340 (ET2451) and 456 (ET4062) showed 16- and 45-fold selectivity, respectively, when comparing α4β7 versus α4β1 cell adhesion assays. In contrast, multimeric compounds based on monomeric Compound No. 340 (ET2451) exhibited 20- to 100-fold selectivity in favor of integrin α4β7, and multimeric compounds based on monomeric Compound No. 456 (ET4062) exhibited no measurable effect on the adhesion of a431-expressing Ramos cells to VCAM (Table 2C).

In Vivo T Lymphocyte Trafficking Analyses

The ability of several integrin alpha-4-beta-7-inhibiting compounds to attenuate the trafficking of integrin alpha-4-beta-7-expressing T lymphocytes was demonstrated in in vivo pharmacodynamics studies in DSS-treated mice. Dextran Sodium Sulfate (DSS) induces chronic colitis in experimental animals when given orally in drinking water for five days followed by no DSS in drinking water. Chronic inflammation is associated with the infiltration of leucocytes from the blood to intestinal tissues. The interaction between integrin α4β7 and MAdCAM-1 on the endothelium of the gut allows adhesion and trafficking of T cells to the gut. The ability of several integrin alpha-4-beta-7-inhibiting nacellins to attenuate the trafficking of integrin alpha-4-beta-7-expressing T lymphocytes was demonstrated in in vivo pharmacodynamics studies in DSS-treated mice.

A study was conducted in which mice were exposed for 5 days to dextran sulfate in their drinking water. On days 6 to 9, compounds or vehicle were administered orally daily. Mesenteric lymph nodes were collected 4 hours following the last dose and assessed. As shown in FIG. 8, Compound No. 517 (ET3764) reduced the detection of integrin □4□7+ T helper memory lymphocytes in the mesenteric lymph nodes (MLN). Compound No. 517, administered at a dose of 80 mg/kg, reduced the number of α4β7+ positive lymphocytes by 60%.

We determined that the level of reduction in α4β7+ T helper memory lymphocytes detected in the mesenteric lymph nodes of DSS treated mice was dependent on the dose of Compound No. 517 administered. FIG. 9 shows the dose-dependent reduction in α4β7+ T cells present in the mesenteric lymph nodes.

We compared the ability of compounds to inhibit the binding of labeled human recombinant MADCAM-1 or VCAM to α4β7-positive or α4β7-negative Th memory cells respectively. Whole blood from a single donor was incubated with compounds and saturated amounts of recombinant ligands. The inhibition of MAdCAM or VCAM binding was measured on T cell subsets using FACS analysis. As shown in FIG. 10, representative multimeric Compound No.s 517, 482, 530 and 534 inhibited MAdCAM-1 binding to primary cells with IC50 values ranging from 87 to 141 nM. The same representative compounds bound to VCAM with lower affinity, with IC50 values ranging from 600 nM to undetectable binding at 4000 nM (FIG. 11).

Although preferred embodiments of the invention have been described herein, it will be understood by those skilled in the art that variations may be made thereto without departing from the spirit of the invention or the scope of the appended claims. All documents disclosed herein, including those in the following reference list, are incorporated by reference.

TABLE 1A Compound No. R¹ R² R³ R⁴ R⁵  1 H H CH2—S—Ph H C(O)—NH-tert-Butyl  2 H H CH2—S—Ph H C(O)—NH-tert-Butyl  3 H H CH2—S—Ph H C(O)—NH-tert-Butyl  4 H H CH₃ H C(O)—NH-tert-Butyl  5 H H CH₃ H C(O)—NH-tert-Butyl  6 H H CH₃ H C(O)—NH-tert-Butyl  7 H H CH₃ H C(O)—NH-tert-Butyl  8 H H CH₃ H C(O)—NH-tert-Butyl  9 H H CH₃ H C(O)—NH-tert-Butyl  10 H H CH₃ H C(O)—NH-tert-Butyl  11 H H CH₃ H C(O)—NH-tert-Butyl  12 H H CH₃ H C(O)—NH-tert-Butyl  13 H H CH₃ H C(O)—NH-tert-Butyl  14 H H CH₃ H C(O)—NH-tert-Butyl  15 H H CH₃ H C(O)—NH-tert-Butyl  16 H H CH₃ H C(O)—NH-tert-Butyl  17 H H CH₃ H C(O)—NH-tert-Butyl  18 H H CH₃ H C(O)—NH-tert-Butyl  19 H H CH₃ H C(O)—NH-tert-Butyl  20 H H CH₃ H C(O)—NH-tert-Butyl  21 H H CH₃ H C(O)—NH-tert-Butyl  22 H H CH₃ H C(O)—NH-tert-Butyl  23 H H CH₃ H C(O)—NH-tert-Butyl  24 H H CH₃ H C(O)—NH-tert-Butyl  25 H H CH₃ H C(O)—NH-tert-Butyl  26 H H CH₃ H C(O)—NH-tert-Butyl  27 H H CH₃ H C(O)—NH-tert-Butyl  28 H H CH₃ H C(O)—NH-tert-Butyl  29 H H CH₃ H C(O)—NH-tert-Butyl  30 H H CH₃ H C(O)—NH-tert-Butyl  31 H H CH₃ H C(O)—NH-tert-Butyl  32 H H CH₃ H C(O)—NH-tert-Butyl  33 H H CH₃ H C(O)—NH-tert-Butyl  34 H H CH₃ H C(O)—NH-tert-Butyl  35 H H CH₃ H C(O)—NH-tert-Butyl  36 H H CH₃ H C(O)—NH-tert-Butyl  37 H H CH₃ H C(O)—NH-tert-Butyl  38 H H CH₃ H C(O)—NH-tert-Butyl  39 H H CH₃ H C(O)—NH-tert-Butyl  40 H H CH₃ H C(O)—NH-tert-Butyl  41 H H CH₃ H C(O)—NH-tert-Butyl  42 H H CH₃ H C(O)—NH-tert-Butyl  43 H CH₃ H H C(O)—NH-tert-Butyl  44 H H CH₃ H C(O)—NH-tert-Butyl  45 H H CH₃ H C(O)—NH-tert-Butyl  46 H H CH₃ H C(O)—NH-tert-Butyl  47 H H CH₃ H C(O)—NH-tert-Butyl  48 H H CH₃ H C(O)—NH-tert-Butyl  49 H H CH₃ H C(O)—NH-tert-Butyl  50 H H CH₃ H C(O)—NH-tert-Butyl  51 H H CH₃ H C(O)—NH-tert-Butyl  52 H H CH₃ H C(O)—NH-tert-Butyl  53 H H CH₃ H C(O)—NH-tert-Butyl  54 H H CH₃ H C(O)—NH-tert-Butyl  55 H H CH₃ H C(O)—NH-tert-Butyl  56 H H CH₃ H C(O)—NH-tert-Butyl  57 H H CH₃ H C(O)—NH-tert-Butyl  58 H H CH₃ H C(O)—NH-tert-Butyl  59 H H CH₃ H C(O)—NH-tert-Butyl  60 H H CH₃ H C(O)—NH-tert-Butyl  61 H H CH₃ H C(O)—NH-tert-Butyl  62 H H CH₃ H C(O)—NH-tert-Butyl  63 H H CH₃ H C(O)—NH-tert-Butyl  64 H H CH₃ H C(O)—NH-tert-Butyl  65 H H CH₃ H C(O)—NH-tert-Butyl  66 H H CH₃ H C(O)—NH-tert-Butyl  67 H H CH₃ H C(O)—NH-tert-Butyl  68 H H CH₃ H C(O)—NH-tert-Butyl  69 H H CH₃ H C(O)—NH-tert-Butyl  70 H H CH₃ H C(O)—NH-tert-Butyl  71 H H CH₃ H C(O)—NH-tert-Butyl  72 H H CH₃ H C(O)—NH-tert-Butyl  73 H H CH₃ H C(O)—NH-tert-Butyl  74 H H CH₃ H C(O)—NH-tert-Butyl  75 H H CH₃ H C(O)—NH-tert-Butyl  76 H H CH₃ H C(O)—NH-tert-Butyl  77 H H CH₃ H C(O)—NH-tert-Butyl  78 H H CH₃ H C(O)—NH-tert-Butyl  79 H H CH₃ H C(O)—NH-tert-Butyl  80 H H CH₃ H C(O)—NH-tert-Butyl  81 H H CH₃ H C(O)—NH-tert-Butyl  82 H H CH₃ H C(O)—NH-tert-Butyl  83 H H CH₃ H C(O)—NH-tert-Butyl  84 H H CH₃ H C(O)—NH-tert-Butyl  85 H H CH₃ H C(O)—NH-tert-Butyl  86 H H CH₃ H C(O)—NH-tert-Butyl  87 H H CH₃ H C(O)—NH-tert-Butyl  88 H H CH₃ H C(O)—NH-tert-Butyl  89 H H CH₃ H C(O)—NH-tert-Butyl  90 H H CH₃ H C(O)—NH-tert-Butyl  91 H H CH₃ H C(O)—NH-tert-Butyl  92 H H CH₃ H C(O)—NH-tert-Butyl  93 H H CH₃ H C(O)—NH-tert-Butyl  94 H H CH₃ H C(O)—NH-tert-Butyl  95 H H CH₃ H C(O)—NH-tert-Butyl  96 H H CH₃ H C(O)—NH-tert-Butyl  97 H H CH₃ H C(O)—NH-tert-Butyl  98 H H CH₃ H C(O)—NH-tert-Butyl  99 H H CH₃ H C(O)—NH-tert-Butyl 100 H H CH₃ H C(O)—NH-tert-Butyl 101 H H CH₃ H C(O)—NH-tert-Butyl 102 H H CH₃ H C(O)—NH-tert-Butyl 103 H CH₃ H H C(O)—NH-tert-Butyl 104 H H CH₃ H C(O)—NH-tert-Butyl 105 H H CH₃ H C(O)—NH-tert-Butyl 106 H H CH₃ H C(O)—NH-tert-Butyl 107 H H CH₃ H C(O)—NH-tert-Butyl 108 H H CH₃ H C(O)—NH-tert-Butyl 109 H H CH₃ H C(O)—NH-tert-Butyl 110 H H CH₃ H C(O)—NH-tert-Butyl 111 H H CH₃ H C(O)—NH-tert-Butyl 112 H H CH₃ H C(O)—NH-tert-Butyl 113 H H CH₃ H C(O)—NH-tert-Butyl 114 H H CH₃ H C(O)—NH-tert-Butyl 115 H H CH₃ H C(O)—NH-tert-Butyl 116 H H CH₃ H C(O)—NH-tert-Butyl 117 H H CH₃ H C(O)—NH-tert-Butyl 118 H H CH₃ H C(O)—NH-tert-Butyl 119 H H CH₃ H C(O)—NH-tert-Butyl 120 H H CH₃ H C(O)—NH-tert-Butyl 121 H H CH₃ H C(O)—NH-tert-Butyl 122 H H CH₃ H C(O)—NH-tert-Butyl 123 H H CH₃ H C(O)—NH-tert-Butyl 124 H CH₃ H H C(O)—NH-tert-Butyl 125 H H CH₃ H C(O)—NH-tert-Butyl 126 H H CH₃ H C(O)—NH-tert-Butyl 127 H H CH₃ H C(O)—NH-tert-Butyl 128 H H CH₃ H C(O)—NH-tert-Butyl 129 H H CH₃ H C(O)—NH-tert-Butyl 130 H H CH₃ H C(O)—NH-tert-Butyl 131 H H CH₃ H C(O)—NH-tert-Butyl 132 H H CH₃ H C(O)—NH-tert-Butyl 133 H H CH₃ H C(O)—NH-tert-Butyl 134 H H CH₃ H C(O)—NH-tert-Butyl 135 H H CH₃ H C(O)—NH-tert-Butyl 136 H H CH₃ H C(O)—NH-tert-Butyl 137 H H CH₃ H C(O)—NH-tert-Butyl 138 H H CH₃ H C(O)—NH-tert-Butyl 139 H H CH₃ H C(O)—NH-tert-Butyl 140 H H CH₃ H C(O)—NH-tert-Butyl 141 H H CH₃ H C(O)—NH-tert-Butyl 142 H CH₃ H C(O)—NH-tert-Butyl H 143 H H CH₃ H C(O)—NH-tert-Butyl 144 H H CH₃ H C(O)—NH-tert-Butyl 145 H H CH₃ H C(O)—NH-tert-Butyl 146 H H CH₃ H C(O)—NH-tert-Butyl 147 H H CH₃ H C(O)—NH-tert-Butyl 148 H H CH₃ H C(O)—NH-tert-Butyl 149 H H CH₃ H C(O)—NH-tert-Butyl 150 H H CH₃ H C(O)—NH-tert-Butyl 151 H H CH₃ H C(O)—NH-tert-Butyl 152 H H CH₃ H C(O)—NH-tert-Butyl 153 H H CH₃ H C(O)—NH-tert-Butyl 154 H H CH₃ H C(O)—NH-tert-Butyl 155 H H CH₃ H C(O)—NH-tert-Butyl 156 H H CH₃ H C(O)—NH-tert-Butyl 157 H H CH₃ H C(O)—NH-tert-Butyl 158 H H CH₃ H C(O)—NH-tert-Butyl 159 H H CH₃ H C(O)—NH-tert-Butyl 160 H H CH₃ H C(O)—NH-tert-Butyl 161 H H CH₃ H C(O)—NH-tert-Butyl 162 H H CH₃ H C(O)—NH-tert-Butyl 163 H H CH₃ H C(O)—NH-tert-Butyl 164 H H CH₃ H C(O)—NH-tert-Butyl 165 H H CH₃ H C(O)—NH-tert-Butyl 166 H H CH₃ H C(O)—NH-tert-Butyl 167 H H CH₃ H C(O)—NH-tert-Butyl 168 H H CH₃ H C(O)—NH-tert-Butyl 169 H H CH₃ H C(O)—NH-tert-Butyl 170 H H CH₃ H C(O)—NH-tert-Butyl 171 H H CH₃ H C(O)—NH-tert-Butyl 172 H H CH₃ H C(O)—NH-tert-Butyl 173 H H CH₃ H C(O)—NH-tert-Butyl 174 H H CH₃ H C(O)—NH-tert-Butyl 175 H H CH₃ H C(O)—NH-tert-Butyl 176 H H CH₃ H C(O)—NH-tert-Butyl 177 H H CH₃ H C(O)—NH-tert-Butyl 178 H H CH₃ H C(O)—NH-tert-Butyl 179 H H CH₃ H C(O)—NH-tert-Butyl 180 H H CH₃ H C(O)—NH-tert-Butyl 181 H H CH₃ H C(O)—NH-tert-Butyl 182 H H CH₃ H C(O)—NH-tert-Butyl 183 H H CH₃ H C(O)—NH-tert-Butyl 184 H H CH₃ H C(O)—NH-tert-Butyl 185 H H CH₃ H C(O)—NH-tert-Butyl 186 H H CH₃ H C(O)—NH-tert-Butyl 187 H H CH₃ H C(O)—NH-tert-Butyl 188 H H CH₃ H C(O)—NH-tert-Butyl 189 H H CH₃ H C(O)—NH-tert-Butyl 190 H H CH₃ H C(O)—NH-tert-Butyl 191 H H CH₃ H C(O)—NH-tert-Butyl 192 H H CH₃ H C(O)—NH-tert-Butyl 193 H H CH₃ H C(O)—NH-tert-Butyl 194 H H CH₃ H C(O)—NH-tert-Butyl 195 H H CH₃ H C(O)—NH-tert-Butyl 196 H H CH₃ H C(O)—NH-tert-Butyl 197 H H CH₃ H C(O)—NH-tert-Butyl 198 H H CH₃ H C(O)—NH-tert-Butyl 199 H H CH₃ H C(O)—NH-tert-Butyl 200 H H CH₃ H C(O)—NH-tert-Butyl 201 H H CH₃ H C(O)—NH-tert-Butyl 202 H H CH₃ H C(O)—NH-tert-Butyl 203 H H CH₃ H C(O)—NH-tert-Butyl 204 H H CH₃ H C(O)—NH-tert-Butyl 205 H H CH₃ H C(O)—NH-tert-Butyl 206 H H CH₃ H C(O)—NH-tert-Butyl 207 H H CH₃ H C(O)—NH-tert-Butyl 208 H H CH₃ H C(O)—NH-tert-Butyl 209 H H CH₃ H C(O)—NH-tert-Butyl 210 H H CH₃ H C(O)—NH-tert-Butyl 211 H H CH₃ H C(O)—NH-tert-Butyl 212 H H CH₃ H C(O)—NH-tert-Butyl 213 H H CH₃ H C(O)—NH-tert-Butyl 214 H H CH₃ H C(O)—NH-tert-Butyl 215 H H CH₃ H C(O)—NH-tert-Butyl 216 H H CH₃ H C(O)—NH-tert-Butyl 217 H H CH₃ H C(O)—NH-tert-Butyl 218 H H CH₃ H C(O)—NH-tert-Butyl 219 H H CH₃ H C(O)—NH-tert-Butyl 220 H H CH₃ H C(O)—NH-tert-Butyl 221 H H CH₃ H C(O)—NH-tert-Butyl 222 H H CH₃ H C(O)—NH-tert-Butyl 223 H H CH₃ H C(O)—NH-tert-Butyl 224 H H CH₃ H C(O)—NH-tert-Butyl 225 H H CH₃ H C(O)—NH-tert-Butyl 226 H H CH₃ H C(O)—NH-tert-Butyl 227 H H CH₃ H C(O)—NH-tert-Butyl 228 H H CH₃ H C(O)—NH-tert-Butyl 229 H H CH₃ H C(O)—NH-tert-Butyl 230 H H CH₃ H C(O)—NH-tert-Butyl 231 H H CH₃ H C(O)—NH-tert-Butyl 232 H H CH₃ H C(O)—NH-tert-Butyl 233 H H CH₃ H C(O)—NH-tert-Butyl 234 H H CH₃ H C(O)—NH-tert-Butyl 235 H H CH₃ H C(O)—NH-tert-Butyl 236 H H CH₃ H C(O)—NH-tert-Butyl 237 H H CH₃ H C(O)—NH-tert-Butyl 238 H H CH₃ H C(O)—NH-tert-Butyl 239 H H CH₃ H C(O)—NH-tert-Butyl 240 H H CH₃ H C(O)—NH-tert-Butyl 241 H H CH₃ H C(O)—NH-tert-Butyl 242 H H CH₃ H C(O)—NH-tert-Butyl 243 H H CH₃ H C(O)—NH-tert-Butyl 244 H H CH₃ H C(O)—NH-tert-Butyl 245 H H CH₃ H C(O)—NH-tert-Butyl 246 H H CH₃ H C(O)—NH-tert-Butyl 247 H H CH₃ H C(O)—NH-tert-Butyl 248 H H CH₃ H C(O)—NH-tert-Butyl 249 H H CH₃ H C(O)—NH-tert-Butyl 250 H H CH₃ H C(O)—NH-tert-Butyl 251 H H CH₃ H C(O)—NH-tert-Butyl 252 H H CH₃ H C(O)—NH-tert-Butyl 253 H H CH₃ H C(O)—NH-tert-Butyl 254 H H CH₃ H C(O)—NH-tert-Butyl 255 H H CH₃ H C(O)—NH-tert-Butyl 256 H H CH₃ H C(O)—NH-tert-Butyl 257 H H CH₃ H C(O)—NH-tert-Butyl 258 H H CH₃ H C(O)—NH-tert-Butyl 259 H H CH₃ H C(O)—NH-tert-Butyl 260 H H CH₃ H C(O)—NH-tert-Butyl 261 H H CH₃ H C(O)—NH-tert-Butyl 262 H H CH₃ H C(O)—NH-tert-Butyl 263 H H CH₃ H C(O)—NH-tert-Butyl 264 H H CH₃ H C(O)—NH-tert-Butyl 265 H H CH₃ H C(O)—NH-tert-Butyl 266 H H CH₃ H C(O)—NH-tert-Butyl 267 H H CH₃ H C(O)—NH-tert-Butyl 268 H H CH₃ H C(O)—NH-tert-Butyl 269 H H CH₃ H C(O)—NH-tert-Butyl 270 H H CH₃ H C(O)—NH-tert-Butyl 271 H H CH₃ H C(O)—NH-tert-Butyl 272 H H CH₃ H C(O)—NH-tert-Butyl 273 H H CH₃ H C(O)—NH-tert-Butyl 274 H H CH₃ H C(O)—NH-tert-Butyl 275 H H CH₃ H C(O)—NH-tert-Butyl 276 H H CH₃ H C(O)—NH-tert-Butyl 277 H H CH₃ H C(O)—NH-tert-Butyl 278 H H CH₃ H C(O)—NH-tert-Butyl 279 H H CH₃ H C(O)—NH-tert-Butyl 280 H H CH₃ H C(O)—NH-tert-Butyl 281 H H CH₃ H C(O)—NH-tert-Butyl 282 H H CH₃ H C(O)—NH-tert-Butyl 283 H H CH₃ H C(O)—NH-tert-Butyl 284 H H CH₃ H C(O)—NH-tert-Butyl 285 H H CH₃ H C(O)—NH-tert-Butyl 286 H H CH₃ H C(O)—NH-tert-Butyl 287 H H CH₃ H C(O)—NH-tert-Butyl 288 H H CH₃ H C(O)—NH-tert-Butyl 289 H H CH₃ H C(O)—NH-tert-Butyl 290 H H CH₃ H C(O)—NH-tert-Butyl 291 H H CH₃ H C(O)—NH-tert-Butyl 292 H H CH₃ H C(O)—NH-tert-Butyl 293 H H CH₃ H C(O)—NH-tert-Butyl 294 H H CH₃ H C(O)—NH-tert-Butyl 295 PRO- PRO- H H C(O)—NH-tert-Butyl 296 H H CH₃ H C(O)—NH-tert-Butyl 297 H H CH₃ H C(O)—NH-tert-Butyl 298 H H CH₃ H C(O)—NH-tert-Butyl 299 H H CH₃ H C(O)—NH-tert-Butyl 300 H H CH₃ H C(O)—NH-tert-Butyl 301 H H CH₃ H C(O)—NH-tert-Butyl 302 H H CH₃ H C(O)—NH-tert-Butyl 303 H H CH₃ H C(O)—NH-tert-Butyl 304 H H CH₃ H C(O)—NH-tert-Butyl 305 H H CH₃ H C(O)—NH-tert-Butyl 306 H H CH₃ H C(O)—NH-tert-Butyl 307 H H CH₃ H C(O)—NH-tert-Butyl 308 H H CH₃ H C(O)—NH-tert-Butyl 309 H H CH₃ H C(O)—NH-tert-Butyl 310 H H CH₃ H C(O)—NH-tert-Butyl 311 H H CH₃ H C(O)—NH-tert-Butyl 312 H H CH₃ H C(O)—NH-tert-Butyl 313 H H CH₃ H C(O)—NH-tert-Butyl 314 H H CH₃ H C(O)—NH-tert-Butyl 315 H H CH₃ H C(O)—NH-tert-Butyl 316 H H CH₃ H C(O)—NH-tert-Butyl 317 H H CH₃ H C(O)—NH-tert-Butyl 318 H H CH₃ H C(O)—NH-tert-Butyl 319 H H CH₃ H C(O)—NH-tert-Butyl 320 H H CH₃ H C(O)—NH-tert-Butyl 321 H H CH₃ H C(O)—NH-tert-Butyl 322 H H CH₃ H C(O)—NH-tert-Butyl 323 H H CH₃ H C(O)—NH-tert-Butyl 324 H H CH₃ H C(O)—NH-tert-Butyl 325 H H CH₃ H C(O)—NH-tert-Butyl 326 H H CH₃ H C(O)—NH-tert-Butyl 327 H H CH₃ H C(O)—NH-tert-Butyl 328 H H CH₃ H C(O)—NH-tert-Butyl 329 H H CH₃ H C(O)—NH-tert-Butyl 330 H H CH₃ H C(O)—NH-tert-Butyl 331 H H CH₃ H C(O)—NH-tert-Butyl 332 H H CH₃ H C(O)—NH-tert-Butyl 333 H H CH₃ H C(O)—NH-tert-Butyl 334 H H CH₃ H C(O)—NH-tert-Butyl 335 H H CH₃ H C(O)—NH-tert-Butyl 336 H H CH₃ H C(O)—NH-tert-Butyl 337 H H CH₃ H C(O)—NH-tert-Butyl 338 H H CH₃ H C(O)—NH-tert-Butyl 339 H H CH₃ H C(O)—NH-tert-Butyl 340 H H CH₃ H C(O)—NH-tert-Butyl 341 H H CH₃ H C(O)—NH-tert-Butyl 342 H H CH₃ H C(O)—NH-tert-Butyl 343 H H CH₃ H C(O)—NH-tert-Butyl 344 H CH₃ H C(O)—NH-tert-Butyl H 345 H H CH₃ H C(O)—NH-tert-Butyl 346 H H CH₃ H C(O)—NH-tert-Butyl 347 H H CH₃ H C(O)—NH-tert-Butyl 348 H H CH₃ H C(O)—NH-tert-Butyl 349 H H CH₃ H C(O)—NH-tert-Butyl 350 H H CH₃ H C(O)—NH-tert-Butyl 351 H H CH₃ C(O)—NH-tert-Butyl H 352 H H CH₃ H C(O)—NH-tert-Butyl 353 H H CH₃ H C(O)—NH-tert-Butyl 354 H H CH₃ H C(O)—NH-tert-Butyl 355 H H CH₃ H C(O)—NH-tert-Butyl 356 H H CH₃ H C(O)—NH-tert-Butyl 357 H H CH₃ H C(O)—NH-tert-Butyl 358 H H CH₃ H C(O)—NH-tert-Butyl 359 H H CH₃ H C(O)—NH-tert-Butyl 360 H H CH₃ H C(O)—NH-tert-Butyl 361 H H CH₃ H C(O)—NH-tert-Butyl 362 H H CH₃ H C(O)—NH-tert-Butyl 363 H H CH₃ H C(O)—NH-tert-Butyl 364 H H CH₃ H C(O)—NH-tert-Butyl 365 H H CH₃ H C(O)—NH-tert-Butyl 366 H H CH₃ H C(O)—NH-tert-Butyl 367 H H CH₃ H C(O)—NH-tert-Butyl 368 H H CH₃ H C(O)—NH-tert-Butyl 369 H H CH₃ H C(O)—NH-tert-Butyl 370 H H CH₃ H C(O)—NH-tert-Butyl 371 H H CH₃ H C(O)—NH-tert-Butyl 372 H H CH₃ H C(O)—NH-tert-Butyl 373 H H CH₃ H C(O)—NH-tert-Butyl 374 H H CH₃ H C(O)—NH-tert-Butyl 375 H H CH₃ H C(O)—NH-tert-Butyl 376 H H CH₃ H C(O)—NH-tert-Butyl 377 H H CH₃ H C(O)—NH-tert-Butyl 378 H H CH₃ H C(O)—NH-tert-Butyl 379 H H CH₃ H C(O)—NH-tert-Butyl 380 H H CH₃ H C(O)—NH-tert-Butyl 381 H H CH₃ H C(O)—NH-tert-Butyl 382 H H CH₃ H C(O)—NH-tert-Butyl 383 H H CH₃ H C(O)—NH-tert-Butyl 384 H H CH₃ H C(O)—NH-tert-Butyl 385 H H CH₃ H C(O)—NH-tert-Butyl 386 H H CH₃ H C(O)—NH-tert-Butyl 387 H H CH₃ H C(O)—NH-tert-Butyl 388 H H CH₃ H C(O)—NH-tert-Butyl 389 H H CH₃ H C(O)—NH-tert-Butyl 456 H H CH₃ H C(O)—NH-tert-Butyl

TABLE 1B Com- Seq. pound ID. No. No. R⁶ R⁷ R⁸ X^(y) X^(z) X¹ X² X³ 1 1 PRO PRO H Y L D V 2 2 PRO PRO H H L D V 3 3 PRO PRO H Y L D T 4 3 PRO PRO H Y L D T 5 4 PRO PRO H F L D T 6 5 PRO PRO H HomoPhe L D T 7 6 PRO PRO H Cha L D T 8 7 PRO PRO H W L D I 9 8 PRO PRO H 1Nal L D T 10 9 PRO PRO H 2Nal L D T 11 10 PRO PRO H W L D Thr(OBn) 12 11 PRO PRO H Bip L D T 13 12 PRO PRO H Tyr(OPh) L D T 14 13 PRO PRO H 1Nal L D I 15 14 PRO PRO H 2Nal L D I 16 15 PRO PRO H 2Nal L D Thr(OBn) 17 16 [(4S)- [(4S)- H W L D T fluoro- fluoro- Pro] Pro] 18 17 PRO PRO H Bip L D Thr(OBn) 19 18 PRO PRO H Tyr(2-tolyl diaryl ether) L D T 20 19 PRO PRO H Tyr(4-CF3 diaryl ether) L D T 21 20 PRO PRO H Tyr(4-methoxy diaryl ether) L D T 22 21 PRO PRO H Tyr(4-fluoro diaryl ether) L D T 23 22 PRO PRO H Tyr(2-methoxy diaryl ether) L D T 24 23 PRO PRO H Tyr(3-methoxy diaryl ether) L D T 25 24 PRO PRO H Tyr(3-fluoro diaryl ether) L D T 26 25 PRO PRO H Tyr(3,4-difluoro diaryl ether) L D T 27 26 PRO PRO H Tyr(3-methyl diaryl ether) L D T 28 27 PRO PRO H Tyr(3,4-dimethyl diaryl ether) L D T 29 28 PRO PRO H Tyr(4-CO2Me diaryl ether) L D T 30 29 PRO PRO H Tyr(3-CO2Me diaryl ether) L D T 31 30 PRO PRO H Tyr(4-CO2H diaryl ether) L D T 32 31 HYP HYP H F L D T 393 31 HYP HYP H F L D T 394 31 HYP HYP H F L D T 395 31 HYP HYP H F L D T 396 31 HYP HYP H F L D T 397 31 HYP HYP H F L D T 33 32 PRO PRO H metaY(Opr) L D T 34 33 PRO PRO H Orn(benzamide) L D Thr(OBn) 35 34 PRO PRO H Orn(acetamide) L D Thr(OBn) 36 35 PRO PRO H Orn(methanesulfonamide) L D Thr(OBn) 37 36 PRO PRO H Orn(ethylcarbamate) L D Thr(OBn) 38 37 PRO PRO H Orn(pentyl amide) L D Thr(OBn) 39 38 PRO PRO H R L D T 40 39 PRO PRO H F L D Thr(OMe) 41 40 PRO PRO H F L D Thr(OEt) 42 41 PRO PRO H dTyr L D T 43 42 PRO PRO H dTic L D T 69 42 PRO PRO H dTic L D T 44 43 HYP HYP H [3-(3′-pyridyl)-Ala] L D T 45 44 [(4R)- [(4R)- H F L D T fluoro- fluoro- Pro] Pro] 46 45 [(4R)- [(4R)- H Bip L D T fluoro- fluoro- Pro] Pro] 47 46 [(4R)- [(4R)- H [3-(3′-pyridyl)-Ala] L D T fluoro- fluoro- Pro] Pro] 48 47 [(4R)- [(4R)- H Y L D T fluoro- fluoro- Pro] Pro] 49 48 [(4S)- [(4S)- H Y L D T fluoro- fluoro- Pro] Pro] 50 49 PRO PRO H dArg L D T 51 50 PRO PRO H dPip L D T 52 51 PRO PRO H [3-(4-thiazolyl)-Ala] L D T 53 52 PRO PRO H Y L D I 54 53 PRO PRO H (4-aza-Phe) L D T 55 54 PRO PRO H Y L D Pen 56 55 PRO PRO H (vinyl-Br-Leu) L D T 57 56 PRO PRO H Hyp(OBn) L D T 58 56 PRO PRO H Hyp(OBn) L D T 59 57 PRO PRO H Dap(Cbz) L D T 60 58 PRO PRO H His(Bn) L D T 61 59 PRO PRO H (4-amino-Phe) L D T 62 60 PRO PRO H (4-aza-dPhe) L D T 63 61 PRO PRO H Hyp L D T 64 62 PRO PRO H dTrp L D T 65 63 PRO PRO H M L D T 66 64 PRO PRO H dMet L D T 67 65 PRO PRO H (4-guanidino-Phe) L D T 68 66 PRO PRO H (3-aza-Phe) L D T 70 67 PRO PRO H (3-aza-dPhe) L D T 71 68 PRO PRO H Nva L D T 72 69 PRO PRO H dNle L D T 73 70 PRO PRO H dLys L D T 74 71 PRO PRO H dPro L D T 75 72 PRO PRO H dOrn L D T 76 73 PRO PRO H (3-benzothienyl-Ala) L D T 77 74 PRO PRO H dTyr(OAllyl) L D T 78 75 PRO PRO H dSer(OBn) L D T 79 76 PRO PRO H [3-(4-thiazolyl)-dAla] L D T 80 77 PRO PRO H (3-benzothienyl-dAla) L D T 81 78 PRO PRO H [3-(2-thienyl)-dAla L D T 82 79 PRO PRO H (4-aminomethyl-Phe) L D T 83 80 PRO PRO H dOrn(dimethyl) L D T 84 81 PRO PRO H (4-amino-dPhe) L D T 85 82 PRO PRO H (4-aminomethyl-dPhe) L D T 86 83 PRO PRO H dTyr(OBn) L D T 87 84 PRO PRO H P L D T 88 85 PRO PRO H cycloLeu L D T 89 86 PRO PRO H Aic L D T 90 87 PRO PRO H Tyr(OAllyl) L D T 91 88 PRO PRO H Chg L D T 92 89 PRO PRO H K L D T 93 90 PRO PRO H (2-aza-dPhe) L D T 94 91 PRO PRO H (2-aza-Phe) L D T 95 92 PRO PRO H [2-(2-pyridyl)-4-thiazolyl-Ala] L D T 96 93 PRO PRO H [2-(3-pyridyl)-4-thiazolyl-Ala] L D T 97 94 PRO PRO H [2-(4-pyridyl)-4-thiazolyl-Ala] L D T 98 95 PRO PRO H dTiq L D T 99 96 PRO PRO H [1-(S)-isoindoline-carboxylic acid] L D T 100 97 PRO PRO H Y dThr L D T 101 98 PRO PRO H Y P L D T 102 99 PRO PRO H Y dPro L D T 124 99 PRO PRO H Y dPro L D T 103 100 PRO PRO H Y Sar L D T 105 100 PRO PRO H Y Sar L D T 104 101 PRO PRO H Y cycloLeu L D T 106 102 PRO PRO H (3-iodo-Phe) Sar L D T 107 103 PRO PRO H (4-iodo-Phe) Sar L D T 108 104 PRO PRO H (3,3-diphenyl-Ala) Sar L D T 109 105 PRO PRO H F dLys L D T 110 106 PRO PRO H Bip dLys L D T 111 107 PRO PRO H [3-(4-thiazolyl)-Ala] dLys L D T 112 108 PRO PRO H (3,3-diphenyl-Ala) dLys L D T 113 109 PRO PRO H Y dLys L D I 114 110 PRO PRO H Y dArg L D T 115 111 PRO PRO H Y dSer L D T 116 112 PRO PRO H Bip Sar L D T 117 113 PRO PRO H 1Nal Sar L D T 118 114 PRO PRO H Y Pip L D T 119 115 PRO PRO H (2-iodo-Phe) Sar L D T 120 116 PRO PRO H 1Nal dLys L D T 121 117 PRO PRO H Y dLys L D MeThr 122 118 PRO PRO H F Sar L D T 123 119 PRO PRO H Y dTic L D T 125 120 PRO PRO H Y dPip L D T 126 121 PRO PRO H F dPro L D T 127 122 PRO PRO H (3,4-dimethoxy-Phe) dPro L D T 128 123 PRO PRO H (3,4,5-trifluoro-Phe) dPro L D T 129 124 PRO PRO H (3,5-dibromo-Tyr) dPro L D T 130 125 PRO PRO H F dPip L D T 131 126 PRO PRO H [3-(4-thiazolyl)-Ala] dPip L D T 132 127 PRO PRO H (4-aminomethyl-Phe) dPip L D T 133 128 PRO PRO H [2-iodo-Phe] dPip L D T 134 129 PRO PRO H (2-phenyl-Phe) dPip L D T 135 130 PRO PRO H [2-(2-methoxy-phenyl)-Phe] dPip L D T 136 131 PRO PRO H [2-(3-methoxy-phenyl)-Phe] dPip L D T 137 132 PRO PRO H [2-(4-methoxy-phenyl)-Phe] dPip L D T 138 133 PRO PRO H Bip dPip L D T 139 134 PRO PRO H Y Hyp L D T 140 135 PRO PRO H Y dHyp L D T 141 136 PRO PRO H Y (cis-dHyp) L D T 142 137 dPRO H dPRO dTyr dPip L D T 143 138 PRO PRO H 1Nal dPip L D T 144 139 PRO PRO H 2Nal dPip L D T 145 140 PRO PRO H (4-aminomethyl-Phe) dTic L D T 146 141 PRO PRO H (3-aminomethyl-Phe) dTic L D T 147 142 PRO PRO H (3-aminomethyl-dPhe) dTic L D T 148 143 PRO PRO H MeTyr dPip L D T 149 144 PRO PRO H Y dPip L D alloThr 150 145 PRO PRO H Y dPip tertbutylAla D T 151 146 PRO PRO H [3-(4-thiazolyl)-Ala] dHyp L D T 152 147 PRO PRO H (4-aminomethyl-Phe) dHyp L D T 153 148 PRO PRO H Y dPip L D I 154 149 PRO PRO H Y dMeLys L D I 155 150 PRO PRO H Y dNle L D T 156 151 PRO PRO H F dHyp L D T 157 152 PRO PRO H Y dMeArg L D T 158 153 PRO PRO H Y G L D T 159 154 PRO PRO H Y A L D T 160 155 PRO PRO H Y dAla L D T 161 156 PRO PRO H M G L D T 162 157 PRO PRO H Tyr(OAllyl) Sar L D T 163 158 PRO PRO H Tyr(OAllyl) G L D T 164 159 PRO PRO H [3-(4-thiazolyl)-Ala] Sar L D T 165 160 PRO PRO H (4-aminomethyl-Phe) G L D T 166 161 PRO PRO H Tyr(OAllyl) dVal L D T 167 162 PRO PRO H Tyr(OAllyl) dSer L D T 168 163 PRO PRO H Tyr(OAllyl) dAla L D T 169 164 PRO PRO H Tyr(OAllyl) P L D T 170 165 PRO PRO H Tyr(OAllyl) dPro L D T 171 166 PRO PRO H [3-(4-thiazolyl)-Ala] dVal L D T 172 167 PRO PRO H [3-(4-thiazolyl)-Ala] dSer L D T 173 168 PRO PRO H [3-(4-thiazolyl)-Ala] dAla L D T 174 169 PRO PRO H [3-(4-thiazolyl)-Ala] P L D T 175 170 PRO PRO H [3-(4-thiazolyl)-Ala] dPro L D T 176 171 PRO PRO H (4-aminomethyl-Phe) P L D T 177 172 PRO PRO H (4-aminomethyl-Phe) dPro L D T 178 173 PRO PRO H cycloLeu P L D T 179 174 PRO PRO H [2-(2-pyridyl)-4-thiazolyl-Ala] Sar L D T 180 175 PRO PRO H [2-(2-pyridyl)-4-thiazolyl-Ala] dPro L D T 181 176 PRO PRO H [2-(3-pyridyl)-4-thiazolyl-Ala] Sar L D T 182 177 PRO PRO H [2-(3-pyridyl)-4-thiazolyl-Ala] dPro L D T 183 178 PRO PRO H [2-(4-pyridyl)-4-thiazolyl-Ala] dPro L D T 184 179 PRO PRO H [3-(2-aminobenzyl-4-thiazolyl)-Ala] Sar L D T 185 180 PRO PRO H [2-(amino-benzyl)-4-thiazolyl-Ala] dPro L D T 186 181 PRO PRO H dTyr dPip L D I 187 182 PRO PRO H (2-aminomethyl-Phe) Aze L D T 188 183 PRO PRO H Y dPip L D Abu 189 184 PRO PRO H (3-aminomethyl-Phe) dTic L D Abu 190 185 PRO PRO H (2,4-dichloro-Phe) dPip L D T 191 186 PRO PRO H (3-phenyl-dPhe) dPip L D T 192 187 PRO PRO H [3-(5-quinolinyl)-dPhe] dPip L D T 193 188 PRO PRO H Y betaHomoLys L D T 194 189 PRO PRO H Y betaHomoPro L D T 195 190 PRO PRO H Y betaHomoLys L D T 196 191 PRO PRO H Y 2Abz L D T 197 192 PRO PRO H F betaHomoLys L D T 198 193 PRO PRO H [3-(4-thiazolyl)-Ala] betaHomoLys L D T 199 194 PRO PRO H (4-aminomethyl-Phe) betaHomoLys L D T 200 195 PRO PRO H Y betaHomoLys L D Thr(OBn) 201 196 PRO PRO H MeTyr dbetaHomoLys L D T 202 197 PRO PRO H 1Nal betaHomoLys L D T 203 198 PRO PRO H 2Nal betaHomoLys L D T 204 199 PRO PRO H Bip betaHomoLys L D T 205 200 PRO PRO H (2-iodo-Phe) betaHomoLys L D T 206 201 PRO PRO H [2-(2,5-dimethyl-isoxazole)-Phe] betaHomoLys L D T 207 202 PRO PRO H (2-phenyl-Phe) betaHomoLys L D T 208 202 PRO PRO H (2-phenyl-Phe) betaHomoLys L D T 209 203 PRO PRO H [(2-piperazinyl-2-Phenyl)-Phe] betaHomoLys L D T 210 204 PRO PRO H Cha betaHomoLys L D T 211 205 PRO PRO H W betaHomoLys L D T 212 206 PRO PRO H dTrp betaHomoLys L D T 213 207 PRO PRO H (3-aminomethyl-Phe) betaHomoLys L D T 214 208 PRO PRO H (4-aminomethyl-dPhe) betaHomoLys L D T 215 209 PRO PRO H (4-aminomethyl-Phe) betaHomoLys L D I 216 210 PRO PRO H Y dbetaHomoLys L D I 217 211 PRO PRO H dArg betaHomoLys L D T 218 212 PRO PRO H (4-aminomethyl-Phe)-reduced betaHomoLys L D T 219 213 PRO PRO H [3-(4-thiazolyl)-Ala] dbetaHomoLys L D I 220 214 PRO PRO H F dbetaHomoLys L D I 221 215 PRO PRO H [3-(4-thiazolyl)-Ala] MebetaHomoLys L D T 222 216 PRO PRO H (4-aminomethyl-Phe) MebetaHomoLys L D T 223 217 PRO PRO H [3-(4-thiazolyl)-Ala] betaHomoLys L D I 224 218 PRO PRO H Tic betaHomoLys L D T 225 219 PRO PRO H dTic betaHomoLys L D T 226 220 PRO PRO H dTic dbetaHomoLys L D T 227 221 PRO PRO H Y betaHomolle L D T 228 222 PRO PRO H (4-aminomethyl-Phe) betaHomoPro L D T 229 223 PRO PRO H Y dbetaHomoPro L D T 230 224 PRO PRO H (4-aminomethyl-Phe) dbetaHomoPro L D T 231 225 PRO PRO H R betaHomoLys L D T 232 226 PRO PRO H F MebetaHomoLys L D T 233 227 PRO PRO H Phe-reduced betaHomoLys L D T 234 228 PRO PRO H (3-aminomethyl-dPhe) betaHomoLys L D T 235 229 PRO PRO H [2-[3-(1-piperazinyl)phenyl]-Phe]- betaHomoLys L D T betaHomoLys 236 230 PRO PRO H [3-(4-thiazolyl)-dAla] betaHomoLys L D T 237 231 PRO PRO H (2-bromo-Phe) betaHomoLys L D T 238 232 PRO PRO H (2-chloro-Phe) betaHomoLys L D T 239 233 PRO PRO H (2-fluoro-Phe) betaHomoLys L D T 240 234 PRO PRO H (2-CF3-Phe) betaHomoLys L D T 241 235 PRO PRO H (2,4-dichloro-Phe) betaHomoLys L D T 242 236 PRO PRO H (2-aminomethyl-Phe) betaHomoLys L D T 243 237 PRO PRO H [2-(4-quinolinyl)-Phe] betaHomoLys L D T 244 238 PRO PRO H [2-(5-quinolinyl)-Phe] betaHomoLys L D T 245 239 PRO PRO H [2-(3-quinolinyl)-Phe] betaHomoLys L D T 246 240 PRO PRO H dhomoPhe betaHomoLys L D T 247 241 PRO PRO H (2-iodo-dPhe) betaHomoLys L D T 248 242 PRO PRO H (2-phenyl-dPhe) betaHomoLys L D T 249 243 PRO PRO H [(2-piperazinyl-2-Phenyl)-dPhe] betaHomoLys L D T 250 244 PRO PRO H Y betaHomoLys L D I 251 245 PRO PRO H Y betaHomoLys L D V 252 246 PRO PRO H dTyr betaHomoLys L D I 253 247 PRO PRO H (4-aminomethyl-dPhe) betaHomoLys L D I 254 248 PRO PRO H (4-aminomethyl-Phe) betaHomoLys L D V 255 249 PRO PRO H (3-iodo-Phe) betaHomoLys L D T 256 250 PRO PRO H (3-phenyl-Phe) betaHomoLys L D T 257 251 PRO PRO H [3-(2-methoxy-phenyl)-Phe] betaHomoLys L D T 258 252 PRO PRO H [3-(2,6-dimethoxy-phenyl)-Phe] betaHomoLys L D T 259 253 PRO PRO H [3-(2-trifluoromethoxy-phenyl)-Phe] betaHomoLys L D T 260 254 PRO PRO H (4-iodo-Phe) betaHomoLys L D T 261 255 PRO PRO H [4-(2-methoxy-phenyl)-Phe] betaHomoLys L D T 262 256 PRO PRO H [4-(2-trifluoromethoxy-phenyl)-Phe] betaHomoLys L D T 263 257 PRO PRO H alphaMePhe betaHomoLys L D T 264 258 PRO PRO H MePhe betaHomoLys L D T 265 259 PRO PRO H [3-(2,6-dimethyl-phenyl)-Phe] betaHomoLys L D T 266 260 PRO PRO H [3-(quinolin-4-yl)-Phe] betaHomoLys L D T 267 261 PRO PRO H [3-(3,4-difluoro-phenyl)-Phe] betaHomoLys L D T 268 262 PRO PRO H [4-(2,6-dimethyl-phenyl)-Phe] betaHomoLys L D T 269 263 PRO PRO H [4-(2-chloro-6-methoxy-phenyl)-Phe] betaHomoLys L D T 270 264 PRO PRO H [3-(4-thiazolyl)-Ala]-reduced betaHomoLys L D T 271 265 PRO PRO H [2-[4-(1-piperazinyl)phenyl]-Phe] betaHomoLys L D T 272 266 PRO PRO H [2-(2,6-dimethylphenyl)-Phe] betaHomoLys L D T 273 267 PRO PRO H [2-(benzothiazol-5-yl)-Phe] betaHomoLys L D T 274 268 PRO PRO H HomoPhe betaHomoLys L D T 275 269 PRO PRO H (piperidine-4-amino-4-carboxylic acid) betaHomoLys L D T 276 270 PRO PRO H [2-(2,5-dimethyl-isoxazole)-dPhe] betaHomoLys L D T 277 271 PRO PRO H dTyr betaHomoLys L D V 278 272 PRO PRO H (4-aminomethyl-dPhe) betaHomoLys L D T 279 273 PRO PRO H [2-(2-chloro-6-methoxyphenyl)-Phe] betaHomoLys L D T 280 274 PRO PRO H 2Igl betaHomoLys L D T 281 275 PRO PRO H d2Igl betaHomoLys L D T 282 276 PRO PRO H Atc betaHomoLys L D T 283 277 PRO PRO H Y betaHomoLys L D allolle 284 278 PRO PRO H dTyr betaHomoLys L D allolle 285 279 PRO PRO H (4-aminomethyl-Phe) betaHomoLys L D allolle 286 280 PRO PRO H [2-[2,5-Bis(trifluoromethyl)phenyl]-Phe] betaHomoLys L D T 287 281 PRO PRO H [2-[2,5-Bis(trifluoromethyl)phenyl]-Phe] betaHomoLys L D T 288 282 PRO PRO H Aic betaHomoLys L D T 289 283 PRO PRO H P betaHomoLys L D T 290 284 PRO PRO H dPro betaHomoLys L D T 291 285 PRO PRO H Pip betaHomoLys L D T 292 286 PRO PRO H [2-(3-Pyridyl)-Phe] betaHomoLys L D T 293 287 PRO PRO H [2-(4-Pyridyl)-Phe] betaHomoLys L D T 294 288 PRO PRO H [2-(3-bromo-2-Pyridyl)-Phe] betaHomoLys L D T 295 289 PRO PRO H Y dbetaHomoLys L D T 296 290 PRO PRO H (N-benzyl-Gly) betaHomoLys L D T 297 291 PRO PRO H [2-(2-bromo-3-Pyridyl)-Phe] betaHomoLys L D T 298 292 PRO PRO H [3-(2-chloro-6-methoxy-phenyl)-Phe] betaHomoLys L D T 299 293 PRO PRO H [3-(benzothiazol-5-yl)-Phe] betaHomoLys L D T 300 294 PRO PRO H (2-aminomethyl-Phe) MebetaHomoLys L D T 301 295 PRO PRO H (2-aminomethyl-dPhe) MebetaHomoLys L D T 302 296 PRO PRO H [3-(4-thiazolyl)-dAla] MebetaHomoLys L D T 303 297 PRO PRO H [2-(2-trifluoromethoxy-phenyl)-dPhe] MebetaHomoLys L D T 304 298 PRO PRO H Tic MebetaHomoLys L D T 305 299 PRO PRO H dTic MebetaHomoLys L D T 306 300 PRO PRO H [2-(5-quinolinyl)-dPhe] betaHomoLys L D T 307 301 PRO PRO H Y betaHomoLys L D alloThr 308 302 PRO PRO H Y MebetaHomoLys L D alloThr 309 303 PRO PRO H MeTyr MebetaHomoLys L D T 310 304 PRO PRO H MeTyr MebetaHomoLys L D alloThr 311 305 PRO PRO H MePhe MebetaHomoLys L D T 312 306 PRO PRO H (2-fluoro-Phe) MebetaHomoLys L D T 313 307 PRO PRO H (2-fluoro-MePhe) MebetaHomoLys L D T 314 308 PRO PRO H (2,4-dichloro-Phe) MebetaHomoLys L D T 315 309 PRO PRO H (2,4-dichloro-MePhe) MebetaHomoLys L D T 316 310 PRO PRO H (2-aminomethyl-MePhe) MebetaHomoLys L D T 317 311 PRO PRO H [3-(2,6-dimethoxy-phenyl)-dPhe] betaHomoLys L D T 318 312 PRO PRO H [3-(4-Quinolinyl)-dPhe] betaHomoLys L D T 319 313 PRO PRO H betaHomoLys Aze L D T 320 314 PRO PRO H (3-phenyl-dPhe) betaHomoLys L D T 321 315 PRO PRO H [3-(2-trifluoromethoxy-phenyl)-dPhe] betaHomoLys L D T 322 316 PRO PRO H [3-(2-methoxy-phenyl)-dPhe] betaHomoLys L D T 323 317 PRO PRO H [2-(5-quinolinyl)-MePhe] MebetaHomoLys L D T 324 318 PRO PRO H F betaHomoNle L D T 325 319 PRO PRO H F MebetaHomoLys(Me)2 L D T 326 320 PRO PRO H MePhe MebetaHomoLys(Me)2 L D T 327 321 PRO PRO H M MebetaHomoLys L D T 328 322 PRO PRO H Igl MebetaHomoLys L D T 329 323 PRO PRO H HomoPhe MebetaHomoLys L D T 330 324 PRO PRO H Hyp(OBn) MebetaHomoLys L D T 331 325 PRO PRO H (1,2-cis-ACHC) MebetaHomoLys L D T 332 326 PRO PRO H MeMet MebetaHomoLys L D T 333 327 PRO PRO H betaHomoLys betaHomoLys L D T 334 328 PRO PRO H BetaHomoPhe MebetaHomoLys L D T 335 329 PRO PRO H betahomoMet MebetaHomoLys L D T 336 330 PRO PRO H Y (3-aminomethyl-4- L D T bromo- benzoic acid) 337 331 PRO PRO H Y [3-aminomethyl-4-(4- L D T aza-phenyl)- benzoic acid] 338 332 PRO PRO H Y [3-aminomethyl-4-(2,5- L D T dimethyl-isoxazole)- benzoic acid] 339 333 PRO PRO H Y [3-aminomethyl-4-(3- L D T aminomethyl-phenyl)- benzoic acid] 340 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]- L D T benzoic acid] 356 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl-4- L D T FITC)phenyl]-benzoic acid] 386 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl-4- L D T AlexaFluor 647)phenyl]-benzoic acid] 390 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]- L D T benzoic acid] 391 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]- L D T benzoic acid] 392 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]- L D T benzoic acid] 341 335 PRO PRO H [3-aminomethyl-4-(4-quinolinyl)-benzoic L D T acid] 342 336 PRO PRO H (3-aminomethyl-4-bromo-benzoic acid) L D T 361 336 PRO PRO H (3-aminomethyl-4-bromo-benzoic acid) L D T 343 337 PRO PRO H [3-aminomethyl-4-(2,5-dimethyl-isoxazole)- L D T benzoic acid] 344 338 dPRO H dPRO [3-aminomethyl-4-(4-pyridyl)-benzoic acid] L D T 345 339 PRO PRO H [3-aminomethyl-(4-methylpyrazole-3-yl)- L D T benzoic acid] 346 340 PRO PRO H [3-aminomethyl-4-(3-quinolinyl)-benzoic L D T acid] 347 341 PRO PRO H [3-aminomethyl-4-(5-quinolinyl)-benzoic L D T acid] 348 342 PRO PRO H [3-aminomethyl-4-[2-(1-piperazinyl)phenyl]- L D T benzoic acid] 349 343 PRO PRO H [3-aminomethyl-4-[3-(1-piperazinyl)phenyl]- L D T benzoic acid] 350 344 PRO PRO H [3-aminomethyl-4-[2-(3-(piperidin-4- L D T ylmethoxy)phenyl]-benzoic acid] 351 345 PRO PRO H [3-aminomethyl-4-(4-pyridyl)-benzoic acid] L D T 352 346 PRO PRO H [3-aminomethyl-4-(4-pyridyl)-benzoic acid] L D Thr(OBn) 353 347 PRO PRO H [3-aminomethyl-4-(4-quinolinyl)-benzoic L D alloThr acid] 354 348 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)phenyl]- L D T benzoic acid] 355 349 PRO PRO H [3-aminomethyl-4-(4-quinolinyl)]-benzoic tertbutylAla D T acid 357 350 PRO PRO H (N-benzyl-3-aminomethyl-benzoic acid) L D T 358 351 PRO PRO H (3-aminomethyl-benzoic acid L D T 359 352 PRO PRO H (3-aminomethyl-5-bromo-benzoic acid) L D T 360 353 PRO PRO H (3-aminomethyl-6-bromo-benzoic acid) L D T 362 354 PRO PRO H [3-aminomethyl-5-(4-aza-phenyl)-benzoic L D T acid] 363 355 PRO PRO H [3-aminomethyl-4-(3-thiophenyl)-benzoic L D T acid] 364 356 PRO PRO H [3-aminomethyl-4-(4-N,N-dimethyl- L D T carboxamide-phenyl)-benzoic acid] 365 357 PRO PRO H [3-aminomethyl-4-(4-aza-phenyl)-benzoic L D T acid] 366 358 PRO PRO H [3-aminomethyl-4-(3-aza-phenyl)-benzoic L D T acid] 367 359 PRO PRO H [3-aminomethyl-4-(4-hydroxy-phenyl)- L D T benzoic acid] 368 360 PRO PRO H [3-aminomethyl-4-[5-(2,4-dimethyl)thiazole]- L D T benzoic acid] 369 361 PRO PRO H [3-aminomethyl-4-(3-N,N-dimethylaniline)- L D T benzoic acid] 370 362 PRO PRO H [3-aminomethyl-4-(2-fluoro-pyridyl)-benzoic L D T acid] 371 363 PRO PRO H [3-aminomethyl-4-(5-pyrimidinyl)-benzoic L D T acid] 372 364 PRO PRO H [3-aminomethyl-4-(3-N,N-dimethyl-diaryl L D T ether)-benzoic acid] 373 365 PRO PRO H [3-aminomethyl-4-(3-CF3-phenyl)-benzoic L D T acid] 374 366 PRO PRO H [3-aminomethyl-4-(2,5-dimethoxy-phenyl)- L D T benzoic acid] 375 367 PRO PRO H [3-aminomethyl-4-[(2,3,4-tri-methoxy)- L D T phenyl]-benzoic acid] 376 368 PRO PRO H [3-aminomethyl-4-(4-carboxy)-phenyl)- L D T benzoic acid] 377 369 PRO PRO H [3-aminomethyl-4-(piperonyl)-benzoic acid] L D T 378 370 PRO PRO H (3-aminomethyl-4-piperidinyl-benzoic acid) L D T 379 371 PRO PRO H (3-aminomethyl-4-morpholinyl-benzoic acid) L D T 380 372 PRO PRO H [3-aminomethyl-4-(N,N-dimethyl)-benzoic L D T acid] 381 373 PRO PRO H [3-aminomethyl-4-(2-aminomethylphenyl)- L D T benzoic acid] 382 374 PRO PRO H [3-aminomethyl-4-(3-aminomethylphenyl)- L D T benzoic acid] 383 375 PRO PRO H [3-aminomethyl-4-(4-aminomethylphenyl)- L D T benzoic acid] 384 376 PRO PRO H [3-aminomethyl-4-(4-quinolinyl)-benzoic L D Abu acid] 385 377 H Nva H [3-aminomethyl-4-(4-quinolinyl)-benzoic L D T acid] 387 378 PRO PRO H (N-methyl-3-aminomethyl-benzoic acid) L D T 388 379 PRO PRO H [N-methyl-3-aminomethyl-4-(4-quinolinyl)- L D T benzoic acid] 389 380 PRO PRO H [2-(5-quinolinyl)-Phe]-reduced betaHomoLys L D T 456 427 PRO PRO H K MebetaHomoLys L D T

TABLE 1C Compound ELISA a4b7 Assay ELISA a4b1 Assay ELISA Assay RPMI8866 Adhesion No. IC₅₀ (mM ) IC₅₀ (mM ) Ratio b1/b7 a4b7/MAdCAM IC50 (mM)  1 0.164 0.162 0.988  2 0.109 0.185 1.697  3 0.192 0.475 2.474 25.000  4 0.129 0.357 2.8 11.782  5 0.087 0.062 0.7 7.916  6 0.103 0.200 1.9  7 0.117 0.190 1.6 23.000  8 0.103 0.096 0.9  9 0.061 0.106 1.7  10 0.052 0.070 1.3  11 0.051 0.094 1.8 3.602  12 0.063 0.113 1.8 8.885  13 0.097 0.171 1.8 19.520  14 0.026 0.025 1.0 2.664  15 0.040 0.026 0.7 3.071  16 0.086 0.053 0.6 1.624  17 0.173 26.92  18 0.120  19 0.114 15.044  20 0.146 8.716  21 0.092 9.466  22 0.100 11.556  23 0.176 0.458 2.6 18.880  24 0.087 0.192 2.2 7.632  25 0.096 0.209 2.2 12.431  26 0.088 0.236 2.7 14.070  27 0.067 0.161 2.4 10.478  28 0.117 0.264 2.3 12.562  29 0.073 0.167 2.3 8.133  30 0.058 0.162 2.8 9.277  31 0.057 0.215 3.7 7.950  32 0.100 0.311 3.1 11.161  33 0.090 0.324 3.6 13.059  34 0.043 0.083 1.9 1.153  35 0.039 0.096 2.5 1.230  36 0.112 0.215 1.9 2.392  37 0.036 0.063 1.8 0.856  38 0.065 0.120 1.9 1.899  39 0.152 0.595 3.9 7.576  40 0.063 0.119 1.9  41 0.042 0.106 2.5  42 0.079 0.232 2.9  43 0.026 0.072 2.8  44 0.083 0.188 2.3  45 0.074 0.238 3.2  46 0.106 0.258 2.4  47 0.061 0.135 2.2 6.777  48 0.094 0.332 3.5 20.686  49 0.137 0.326 2.4 17.374  50 0.023 0.290 12.6 3.709  51 0.031 0.102 3.3  52 0.075 0.367 4.9 14.719  53 0.182 21.956  54 0.190 23.916  55 0.113 0.119 1.1  56 0.058 0.200 3.5 4.203  57 0.059 0.148 2.5  58 0.156 0.445 2.9  59 0.197 0.610 3.1  60 0.066 0.214 3.3 6.554  61 0.063 0.223 3.6  62 0.027 0.115 4.3 2.548  63 0.107 0.251 2.3  64 0.046 0.268 5.8 5.367  65 0.005 0.095 18.1 1.033  66 0.093 0.326 3.5 6.348  67 0.075 0.341 4.5 5.093  68 0.067 0.280 4.2 4.158  69 0.022 0.060 2.7 2.646  70 0.035 0.099 2.9 1.163  71 0.184 0.816 4.4  72 0.151 0.409 2.7 7.284  73 0.144 1.247 8.6 17.304  74 0.100 0.763 7.6 15.503  75 0.171 1.209 7.1 13.166  76 0.114 0.466 4.1 6.267  77 0.036 0.185 5.1 5.633  78 0.069 0.272 3.9 6.479  79 0.110 0.552 5.0 13.217  80 0.053 0.556 10.6 3.599  81 0.054 0.241 4.5 5.405  82 0.073 0.213 2.9 5.716  83 0.179 1.226 6.9 32.316  84 0.035 0.218 6.2 6.143  85 0.052 0.206 3.9 4.229  86 0.050 0.167 3.3 4.074  87 0.019 0.269 14.1  88 0.011 0.166 14.9  89 0.016 0.232 14.4  90 0.009 0.317 35.0  91 0.126 1.824 14.5  92 0.053 1.063 19.9  93 0.078 0.311 4.0 6.009  94 0.080 0.250 3.1 9.484  95 0.125 0.303 2.4  96 0.138 0.321 2.3  97 0.124 0.311 2.5  98 0.021 0.058 2.7  99 0.057 0.154 2.7 100 0.132 0.453 3.4 4.446 101 0.129 0.609 4.7 16.092 102 0.021 0.136 6.6 1.464 103 0.108 1.631 15.1 104 0.120 0.506 4.2 105 0.110 1.734 15.8 9.731 106 0.059 1.109 18.7 107 0.150 2.390 16.0 108 0.077 0.814 10.5 13.867 109 0.133 3.312 24.9 15.287 110 0.185 3.923 21.3 21.753 111 0.100 3.923 39.3 12.926 112 0.138 3.008 21.7 17.420 113 0.052 0.709 13.7 7.634 114 0.083 1.889 22.8 6.866 115 0.125 1.121 9.0 15.436 116 0.166 1.385 8.4 117 0.158 1.381 8.7 118 0.112 0.132 1.2 14.202 119 0.079 1.688 21.5 14.057 120 0.157 3.000 19.1 121 0.192 2.187 11.4 122 0.090 1.666 18.6 16.615 123 0.007 0.019 2.5 1.138 124 0.013 0.104 8.3 1.172 125 0.025 0.458 18.4 1.925 126 0.024 0.135 5.6 1.232 127 0.025 0.196 7.8 128 0.026 0.296 11.4 129 0.065 0.636 9.7 130 0.022 0.125 5.6 1.327 131 0.026 0.080 3.1 132 0.029 0.309 10.8 3.626 133 0.015 0.080 5.3 134 0.023 0.178 7.6 135 0.024 0.119 4.9 136 0.032 0.209 6.6 137 0.033 0.254 7.8 138 0.024 0.118 5.0 139 0.100 0.073 0.7 140 0.053 0.512 9.6 141 0.019 0.036 2.0 142 0.164 0.084 0.5 143 0.033 0.068 2.1 144 0.043 0.027 0.6 6.083 145 0.023 0.045 2.0 3.268 146 0.016 0.012 0.7 0.672 147 0.052 0.039 0.8 148 0.086 0.105 1.2 149 0.046 0.546 12.0 12.600 150 0.054 0.447 8.2 151 0.053 0.218 4.1 152 0.102 1.347 13.2 153 0.006 0.017 2.8 0.125 154 0.117 2.664 22.8 155 0.054 1.085 20.3 156 0.019 0.258 13.3 1.412 157 0.067 3.707 55.3 158 0.110 1.537 14.0 15.746 159 0.053 0.467 8.9 41.275 160 0.141 1.349 9.5 8.794 161 0.135 2.035 15.1 6.662 162 0.107 1.875 17.5 16.696 163 0.126 1.389 11.0 22.489 164 0.127 3.288 25.8 30.192 165 0.128 2.918 22.8 30.337 166 0.179 1.382 7.7 167 0.147 1.997 13.6 168 0.077 1.051 13.6 17.847 169 0.176 0.488 2.8 170 0.013 0.104 8.0 1.033 171 0.128 0.658 5.1 14.357 172 0.096 1.030 10.7 9.922 173 0.054 0.719 13.4 12.042 174 0.160 0.619 3.9 175 0.018 0.130 7.2 0.986 176 0.189 1.202 6.3 177 0.019 0.463 24.0 2.853 178 0.027 0.113 4.1 2.710 179 0.174 2.656 15.2 180 0.013 0.068 5.1 0.841 181 0.180 2.272 12.6 182 0.017 0.083 5.0 1.128 183 0.014 0.105 7.5 1.070 184 0.099 0.953 9.6 185 0.018 0.095 5.4 0.662 186 0.062 0.027 0.4 187 0.083 0.404 4.9 188 0.027 0.189 7.0 7.308 189 0.018 0.019 1.0 2.251 190 0.021 0.145 7.0 2.470 191 0.083 4.020 48.4 192 0.118 6.823 57.8 37.800 193 0.092 0.303 3.3 5.621 194 0.038 0.207 5.4 4.617 195 0.049 1.917 38.9 7.931 196 0.158 0.275 1.7 197 0.044 1.327 30.2 7.441 198 0.041 1.223 29.9 5.089 199 0.069 3.138 45.2 19.350 200 0.134 0.352 2.6 201 0.061 0.695 11.4 202 0.086 0.680 8.0 203 0.055 0.534 9.8 204 0.063 0.429 6.8 205 0.047 1.517 32.2 2.231 206 0.046 2.890 63.0 27.621 207 0.025 0.460 18.5 208 0.019 0.522 28.1 4.679 209 0.035 1.977 56.9 16.508 210 0.072 1.148 16.0 211 0.060 2.511 42.2 8.101 212 0.068 2.190 32.1 213 0.055 2.247 41.2 10.605 214 0.069 4.222 60.8 72.055 215 0.033 0.413 12.4 216 0.123 2.509 20.4 217 0.034 1.088 31.8 218 0.190 3.135 16.5 219 0.147 3.253 22.1 220 0.096 1.740 18.2 221 0.015 0.165 11.1 0.248 222 0.013 0.212 16.1 0.325 223 0.015 0.122 8.2 0.549 224 0.055 2.978 53.9 10.962 225 0.099 4.523 45.6 18.130 226 0.094 10.797 115.0 4.076 227 0.034 0.047 1.4 1.491 228 0.034 0.503 14.7 229 0.058 0.075 1.3 230 0.120 0.131 1.1 231 0.031 0.993 32.0 232 0.012 0.110 8.9 0.353 233 0.094 3.861 41.0 19.372 234 0.099 3.203 32.3 235 0.025 1.553 62.6 4.614 236 0.060 6.203 104.2 7.320 237 0.020 0.870 43.9 5.131 238 0.025 1.049 42.3 8.425 239 0.020 0.641 32.3 4.407 240 0.027 0.905 33.2 12.040 241 0.031 3.207 103.4 6.006 242 0.067 5.307 79.0 8.335 243 0.026 0.767 29.4 2.007 244 0.016 0.753 46.7 0.719 245 0.024 0.414 17.5 3.067 246 0.120 17.702 147.1 247 0.035 4.614 132.8 15.134 248 0.045 3.088 69.2 16.371 249 0.045 4.233 94.8 23.107 250 0.017 0.150 8.7 0.401 251 0.024 0.349 14.8 1.386 252 0.032 0.390 12.1 2.408 253 0.069 1.087 15.6 254 0.055 1.803 33.0 255 0.043 3.024 69.7 256 0.072 3.246 45.1 9.562 257 0.058 1.604 27.5 258 0.056 1.584 28.4 259 0.058 5.995 102.8 4.279 260 0.165 9.562 58.1 261 0.096 23.155 241.0 16.926 262 0.080 3.740 47.0 263 0.102 2.345 23.1 264 0.117 5.560 47.5 265 0.039 1.818 46.2 266 0.037 1.206 33.0 11.641 267 0.044 1.936 44.1 20.440 268 0.076 1.868 24.6 269 0.056 1.764 31.6 270 0.160 17.562 109.8 18.900 271 0.033 1.151 34.6 272 0.041 2.383 58.1 273 0.012 0.303 24.6 1.730 274 0.026 0.454 17.5 7.938 275 0.101 0.779 7.7 276 0.134 14.235 106.2 277 0.052 0.357 6.9 278 0.104 1.062 10.2 279 0.100 5.847 58.2 280 0.010 0.400 39.7 2.150 281 0.144 3.161 21.9 282 0.119 0.626 5.2 283 0.128 1.495 11.7 284 0.046 0.228 5.0 285 0.089 0.553 6.2 286 0.064 5.236 81.9 287 0.084 3.553 42.1 288 0.136 1.664 12.2 289 0.038 0.349 9.3 1.242 290 0.067 1.894 28.4 291 0.035 0.777 22.4 8.742 292 0.030 0.374 12.4 293 0.019 0.198 10.6 4.008 294 0.045 0.937 20.7 295 0.094 20.950 222.7 18.900 296 0.155 14.698 94.8 297 0.037 0.786 21.3 298 0.076 4.349 57.2 299 0.002 0.090 41.5 0.556 300 0.022 0.225 10.4 0.672 301 0.018 0.846 47.6 1.020 302 0.012 0.598 51.6 1.764 303 0.020 0.497 24.8 1.662 304 0.015 0.293 19.0 0.191 305 0.008 0.221 26.6 3.533 306 0.104 2.763 26.5 307 0.091 4.343 47.8 308 0.039 0.480 12.3 1.982 309 0.008 0.023 3.0 0.126 310 0.017 0.300 17.6 0.434 311 0.007 0.198 27.6 0.158 312 0.011 0.145 13.4 0.273 313 0.011 0.206 19.2 0.210 314 0.011 0.138 12.8 0.305 315 0.013 0.312 24.9 0.431 316 0.022 0.349 16.2 0.690 317 0.047 0.685 14.5 9.408 318 0.091 1.513 16.6 319 0.065 0.309 4.8 320 0.163 0.127 0.8 321 0.101 7.368 72.7 322 0.093 4.166 44.7 323 0.025 0.297 11.8 1.056 324 0.110 1.058 9.6 11.844 325 0.020 0.170 8.6 0.714 326 0.017 0.476 28.4 0.280 327 0.010 0.128 13.2 0.308 328 0.010 0.234 24.1 0.368 329 0.005 0.050 10.6 0.326 330 0.005 0.179 32.9 0.185 331 0.016 0.093 6.0 0.399 332 0.010 0.120 12.5 0.140 333 0.046 0.757 16.5 12.922 334 5.061 335 4.956 336 0.162 0.917 5.6 337 0.061 0.177 2.9 338 0.041 0.177 4.4 339 0.051 0.299 5.8 340 0.019 0.048 2.5 0.263 341 0.012 0.026 2.1 0.306 342 0.041 0.139 3.4 343 0.018 0.029 1.6 0.269 344 0.052 0.107 2.1 345 0.039 0.052 1.3 346 0.028 0.011 0.4 0.580 347 0.023 0.030 1.3 348 0.027 0.041 1.5 349 0.023 0.043 1.9 0.479 350 0.027 0.055 2.0 351 0.160 0.184 1.2 352 0.024 0.005 0.2 0.070 353 0.031 0.103 3.3 354 0.050 0.175 3.5 355 0.048 0.069 1.4 356 0.017 0.027 1.6 357 0.102 0.406 4.0 358 0.127 1.108 8.7 34.923 359 0.053 0.450 8.5 7.880 360 0.125 0.779 6.2 18.937 361 0.049 0.288 5.9 2.843 362 0.043 0.238 5.6 363 0.022 0.105 4.8 1.571 364 0.018 0.074 4.0 0.602 365 0.017 0.064 3.7 0.638 366 0.023 0.059 2.6 0.384 367 0.018 0.053 3.0 0.535 368 0.010 0.024 2.4 0.342 369 0.024 0.069 2.9 0.974 370 0.015 0.047 3.1 0.661 371 0.016 0.055 3.4 0.482 372 0.024 0.104 4.3 2.133 373 0.018 0.074 4.1 0.879 374 0.018 0.081 4.5 1.246 375 0.015 0.067 4.5 1.164 376 0.019 0.078 4.1 1.135 377 0.013 0.045 3.6 0.839 378 0.042 0.182 4.3 379 0.033 0.161 4.9 380 0.041 0.217 5.3 381 0.010 0.010 1.1 0.323 382 0.012 0.025 2.0 383 0.006 0.017 2.7 0.403 384 0.020 0.049 2.5 2.260 385 0.039 0.023 0.6 1.548 386 0.044 0.034 0.8 2.604 387 0.063 6.133 96.7 16.142 388 0.009 0.102 12.0 0.336 389 0.042 0.234 5.535 6.664 456 0.196

TABLE 1C′ RPMI8866 Ramos ELISA VCAM MADCAM FACS Adhesion Adhesion ELISA a4b7 ELISA a4b1 Assay FACS Compound a4b7 Th mem a4b7/MAdCAM a4b1/VCAM Ratio Assay IC₅₀ Assay IC50 Ratio a4+b7− Th No. (nM) IC50 (nM) IC₅₀ (nM) Ramos/RPMI (nM) (nM) b1/b7 mem (nM) 32 11161 132 3626 146 672 340 Unclear 175 2767 16 456 1000 199 8925 45 14 21 1.5

TABLE 1X Compound LC-MS Experimental No. (m/z) Protocol 1 A, E, Fb, I, M 2 A, E, Fb, I, M 3 A, E, Fb, I, M 4 A, E, Fa, Jb, I, M 4 A, E, Fa, Jb, I, M 5 A, E, Fa, Jb, I, M 5 A, E, Fa, Jb, I, M 6 A, E, Fa, Jb, I, M 7 A, E, Fa, Jb, I, M 8 A, E, Fa, Jb, I, M 9 780.4 A, E, Fa, Jb, I, M 10 780.4 A, E, Fa, Jb, I, M 11 859.4 A, E, Fa, Jb, I, M 12 806.4 A, E, Fa, Jb, I, M 13 A, E, Fa, Jb, H, I, M 14 792.4 A, E, Fa, Jb, I, M 15 792.4 A, E, Fa, Jb, I, M 16 A, E, Fa, Jb, I, M 17 A, E, Fa, Jb, I, M 18 A, E, Fa, Jb, I, M 19 A, E, Fa, Jb, H, I, M 20 A, E, Fa, Jb, H, I, M 21 A, E, Fa, Jb, H, I, M 22 A, E, Fa, Jb, H, I, M 23 A, E, Fa, Jb, H, I, M 24 A, E, Fa, Jb, H, I, M 25 A, E, Fa, Jb, H, I, M 26 A, E, Fa, Jb, H, I, M 27 836.4 A, E, Fa, Jb, H, I, M 28 A, E, Fa, Jb, H, I, M 29 880.4 A, E, Fa, Jb, H, I, M 30 880.4 A, E, Fa, Jb, H, I, M 31 866.4 A, E, Fa, Jb, H, I, M 32 A, E, Fa, Jb, I, M 33 A, E, Fa, Jb, H, I, M 34 891.4 A, E, Fa, Jb, K, I, M 35 829.4 A, E, Fa, Jb, K, I, M 36 A, E, Fa, Jb, K, I, M 37 859.4 A, E, Fa, Jb, K, I, M 38 871.4 A, E, Fa, Jb, K, I, M 39 A, E, Fa, Jb, I, M 40 744.4 A, E, Fa, Jb, I, M 41 758.4 A, E, Fa, Jb, I, M 42 746.4 A, D, I, M 43 742.4 A, D, I, M 44 A, E, Fa, Jb, I, M 45 748.4 A, E, Fa, Jb, I, M 46 A, E, Fa, Jb, I, M 47 749.3 A, E, Fa, Jb, I, M 48 A, E, Fa, Jb, I, M 49 A, E, Fa, Jb, I, M 50 739.4 A, D, I, M 51 A, D, I, M 52 A, D, I, M 53 A, D, I, M 54 A, D, I, M 55 A, D, I, M 56 A, D, I, M 57 A, D, I, M 58 A, D, I, M 59 A, D, I, M 60 A, D, I, M 61 A, D, I, M 62 A, D, I, M 63 A, D, I, M 64 A, D, I, M 65 A, D, I, M 66 A, D, I, M 67 A, D, I, M 68 A, D, I, M 69 A, D, I, M 70 A, D, I, M 71 A, D, I, M 72 A, D, I, M 73 A, D, I, M 74 A, D, I, M 75 A, D, I, M 76 A, D, I, M 77 A, D, I, M 78 A, D, I, M 79 A, D, I, M 80 A, D, I, M 81 A, D, I, M 82 A, D, I, M 83 A, D, I, M 84 A, D, I, M 85 A, D, I, M 86 A, D, I, M 87 A, D, I, M 88 A, D, I, M 89 A, D, I, M 90 A, D, I, M 91 A, D, I, M 92 A, D, I, M 93 A, D, I, M 94 A, D, I, M 95 A, D, G, I, M 96 A, D, G, I, M 97 A, D, G, I, M 98 A, D, I, M 99 A, D, I, M 100 847.4 A, D, I, M 101 843.6 A, D, I, M 102 843.6 A, D, I, M 103 817.4 A, D, I, M 104 857.4 A, D, I, M 105 817.4 A, D, I, M 106 927.2 A, D, I, M 107 927.2 A, D, I, M 108 877.4 A, D, I, M 109 858.4 A, D, I, M 110 934.4 A, D, I, M 111 865.4 A, D, I, M 112 934.4 A, D, I, M 113 886.4 A, D, I, M 114 902.4 A, D, I, M 115 833.4 A, D, I, M 116 877.4 A, D, I, M 117 851.4 A, D, I, M 118 857.4 A, D, I, M 119 927.2 A, D, I, M 120 908.4 A, D, I, M 121 888.4 A, D, I, M 122 801.4 A, D, I, M 123 905.4 A, D, I, M 124 843.4 A, D, I, M 125 857.4 A, D, I, M 125 857.4 A, D, I, M 126 827.4 A, D, I, M 127 887.4 A, D, I, M 128 881.4 A, D, I, M 129 1001.2 A, D, I, M 130 A, D, I, M 131 A, D, I, M 132 A, D, I, M 133 A, D, I, M 134 A, D, G, I, M 135 A, D, G, I, M 136 A, D, G, I, M 137 A, D, G, I, M 138 A, D, I, M 139 A, D, I, M 140 A, D, I, M 141 A, D, I, M 142 A, D, I, M 143 891.1 A, D, I, M 144 891.1 A, D, I, M 145 918.1 A, D, I, M 146 918.1 A, D, I, M 147 918.1 A, D, I, M 148 871.1 A, D, I, M 149 857.1 A, D, I, M 150 871.1 A, D, I, M 151 850.0 A, D, I, M 152 872.1 A, D, I, M 153 869.2 A, D, I, M 154 900.2 A, D, I, M 155 859.4 A, D, I, M 156 843.4 A, D, I, M 157 916.2 A, D, I, M 158 A, D, I, M 159 A, D, I, M 160 A, D, I, M 161 A, D, I, M 162 A, D, I, M 163 A, D, I, M 164 A, D, I, M 165 A, D, I, M 166 A, D, I, M 167 A, D, I, M 168 A, D, I, M 169 A, D, I, M 170 A, D, I, M 171 A, D, I, M 172 A, D, I, M 173 A, D, I, M 174 A, D, I, M 175 A, D, I, M 176 A, D, I, M 177 A, D, I, M 178 A, D, I, M 179 A, D, G, I, M 180 A, D, G, I, M 181 A, D, G, I, M 182 A, D, G, I, M 183 A, D, G, I, M 184 A, D, G, I, M 185 A, D, G, I, M 186 869.3 A, D, I, M 187 842.4 A, D, I, M 188 842.1 A, D, I, M 189 902.1 A, D, I, M 190 909.1 A, D, I, M 191 948.4 A, D, G, I, M 192 968.4 A, D, G, I, M 193 861.4 A, D, I, M 194 857.4 A, D, I, M 195 888.4 A, D, I, M 196 702.4 A, D, I, M 197 A, D, I, M 198 A, D, I, M 199 A, D, I, M 200 A, D, I, M 201 A, D, I, M 202 A, D, I, M 203 A, D, I, M 204 A, D, I, M 205 A, D, I, M 206 967.2 A, D, G, I, M 207 948.2 A, D, G, I, M 208 948.2 A, D, G, I, M 209 A, D, G, I, M 210 A, D, I, M 211 A, D, I, M 212 A, D, I, M 213 A, D, I, M 214 A, D, I, M 215 A, D, I, M 216 900.4 A, D, I, M 217 A, D, I, M 218 A, C, D, I, M 219 A, D, I, M 220 A, D, I, M 221 893.1 A, B, D, I, M 222 915.2 A, B, D, I, M 223 891.1 A, D, I, M 224 A, D, I, M 225 A, D, I, M 226 A, D, I, M 227 A, D, I, M 228 A, D, I, M 229 857.4 A, D, I, M 230 870.4 A, D, I, M 231 A, D, I, M 232 886.2 A, B, D, I, M 232 886.2 A, B, D, I, M 233 A, C, D, I, M 234 A, D, I, M 235 A, D, G, I, M 236 A, D, I, M 237 952.0 A, D, I, M 238 A, D, I, M 239 890.2 A, D, I, M 240 A, D, I, M 241 A, D, I, M 242 A, D, I, M 243 A, D, G, I, M 244 999.3 A, D, G, I, M 245 A, D, G, I, M 246 A, D, I, M 247 A, D, I, M 248 A, D, G, I, M 249 A, D, G, I, M 250 900.2 A, D, I, M 251 886.1 A, D, I, M 252 A, D, I, M 253 A, D, I, M 254 A, D, I, M 255 998.4 A, D, I, M 256 948.6 A, D, G, I, M 257 978.6 A, D, G, I, M 258 1008.6 A, D, G, I, M 259 1032.6 A, D, G, I, M 260 998.4 A, D, I, M 261 978.4 A, D, G, I, M 262 1032.4 A, D, G, I, M 263 886.6 A, D, I, M 264 886.6 A, D, I, M 265 976.6 A, D, G, I, M 266 1000.6 A, D, G, I, M 267 984.5 A, D, G, I, M 268 976.6 A, D, G, I, M 269 1012.6 A, D, G, I, M 270 A, C, D, G, I, M 270 A, C, D, I, M 271 A, D, G, I, M 272 A, D, G, I, M 273 1005.3 A, D, G, I, M 274 A, D, I, M 275 A, D, I, M 276 A, D, G, I, M 277 A, D, I, M 278 A, D, I, M 279 A, D, I, M 280 898.1 A, D, I, M 281 A, D, I, M 282 A, D, I, M 283 A, D, I, M 284 A, D, I, M 285 A, D, I, M 286 A, D, G, I, M 287 A, D, G, I, M 288 A, D, I, M 289 A, D, I, M 290 A, D, I, M 291 A, D, I, M 292 A, D, G, I, M 293 949.2 A, D, G, I, M 294 A, D, G, I, M 295 A, D, I, M 296 A, D, I, M 297 A, D, G, I, M 298 1012.6 A, D, G, I, M 299 1005.4 A, D, G, I, M 300 915.2 A, B, D, I, M 301 915.2 A, B, D, I, M 302 893.2 A, B, D, I, M 303 1047.3 A, B, D, I, M 304 898.2 A, B, D, I, M 305 898.2 A, B, D, I, M 306 A, D, G, I, M 307 A, D, I, M 308 A, B, D, I, M 309 916.2 A, B, D, I, M 310 916.2 A, B, D, I, M 311 900.2 A, B, D, I, M 312 904.2 A, B, D, I, M 313 918.2 A, B, D, I, M 314 954.1 A, B, D, I, M 315 968.1 A, B, D, I, M 316 929.2 A, B, D, I, M 317 A, D, G, I, M 318 999.4 A, D, G, I, M 319 808.6 A, D, I, M 320 917.4 A, D, G, I, M 321 1032.4 A, D, G, I, M 322 978.5 A, D, G, I, M 323 A, B, D, G, I, M 324 A, D, I, M 325 914.3 A, B, D, I, M 326 928.2 A, B, D, I, M 327 870.2 A, B, D, I, M 328 912.2 A, B, D, I, M 329 900.4 A, B, D, I, M 330 942.5 A, B, D, I, M 331 852.5 A, B, D, I, M 332 884.6 A, B, D, I, M 333 867.2 A, D, I, M 334 900.4 A, B, D, I, M 335 884.5 A, B, D, I, M 336 A, D, I, M 337 A, D, G, I, M 338 A, D, G, I, M 339 A, D, G, I, M 340 A, D, G, I, M 341 A, D, G, I, M 341 A, D, G, I, M 342 A, D, I, M 343 A, D, G, I, M 344 A, D, G, I, M 345 A, D, G, I, M 346 A, D, G, I, M 347 A, D, G, I, M 348 A, D, G, I, M 349 A, D, G, I, M 350 A, D, G, I, M 351 A, D, G, I, M 352 A, D, G, I, M 353 A, D, G, I, M 354 A, D, G, I, M 355 A, D, G, I, M 356 A, D, G, I, M, I, M 357 A, D, I, M 358 A, D, I, M 359 A, D, I, M 360 A, D, I, M 361 A, D, I, M 362 A, D, G, I, M 363 A, D, G, I, M 364 A, D, G, I, M 365 A, D, G, I, M 366 A, D, G, I, M 367 A, D, G, I, M 368 A, D, G, I, M 369 A, D, G, I, M 370 A, D, G, I, M 371 A, D, G, I, M 372 A, D, G, I, M 373 A, D, G, I, M 374 A, D, G, I, M 375 A, D, G, I, M 376 A, D, G, I, M 377 A, D, G, I, M 378 A, D, G, I, M 379 A, D, G, I, M 380 A, D, G, I, M 381 A, D, G, I, M 382 A, D, G, I, M 383 A, D, G, I, M 384 A, D, G, I, M 385 A, D, G, I, M 386 A, D, G, I, M 387 A, B, D, I, M 388 A, B, D, G, I, M 389 985.2 A, C, D, G, I, M 398 A, D, I, M 399 A, D, I, M 400 A, D, I, M 401 A, D, I, M 402 A, D, I, M 403 A, D, I, M 404 A, D, I, M 405 A, E, Fa, Jb, I, M 406 A, E, Fa, Jb, I, M 407 A, D, I, M 408 A, D, I, M 409 A, D, I, M 410 A, D, I, M 411 A, E, Fa, Jb, I, M 412 A, D, I, M 413 A, E, Fa, Jb, I, M 414 A, E, Fa, Jb, I, M 415 A, D, I, M 416 A, D, I, M 417 A, D, I, M 418 A, D, I, M 419 A, D, I, M 420 A, D, I, M 421 A, D, I, M 422 A, D, I, M 423 A, D, I, M 424 A, D, I, M 425 A, D, I, M 426 A, D, I, M 427 A, D, I, M 428 A, D, I, M 429 A, D, I, M 430 A, D, I, M 431 A, D, I, M 432 A, D, I, M 433 A, D, G, I, M 434 A, D, G, I, M 435 A, D, G, I, M 436 A, D, G, I, M 437 A, D, G, I, M 438 A, D, G, I, M 439 A, D, I, M 440 A, D, I, M 441 A, D, I, M 442 A, B, D, I, M 443 A, D, I, M 444 A, D, I, M 445 A, D, I, M 446 A, D, I, M 447 A, D, I, M 448 A, D, I, M 449 A, D, I, M 450 A, D, I, M 451 A, C, D, I, M 452 A, C, D, I, M 453 A, C, D, I, M 454 A, C, D, I, M 455 A, C, D, I, M

TABLE 2A Compound No. R¹ R² R³ R⁴ R⁵ 390 H H CH₃ H C(O)—NH-tert-Butyl 391 H H CH₃ H C(O)—NH-tert-Butyl 392 H H CH₃ H C(O)—NH-tert-Butyl 393 H H CH₃ H C(O)—NH-tert-Butyl 394 H H CH₃ H C(O)—NH-tert-Butyl 395 H H CH₃ H C(O)—NH-tert-Butyl 396 H H CH₃ H C(O)—NH-tert-Butyl 397 H H CH₃ H C(O)—NH-tert-Butyl 457 H H CH₃ H C(O)—NH-tert-Butyl 458 H H CH₃ H C(O)—NH-tert-Butyl 459 H H CH₃ H C(O)—NH-tert-Butyl 460 H H CH₃ H C(O)—NH-tert-Butyl 461 H H CH₃ H C(O)—NH-tert-Butyl 462 H H CH₃ H C(O)—NH-tert-Butyl 463 H H CH₃ H C(O)—NH-tert-Butyl 464 H H CH₃ H C(O)—NH-tert-Butyl 465 H H CH₃ H C(O)—NH-tert-Butyl 466 H H CH₃ H C(O)—NH-tert-Butyl 467 H H CH₃ H C(O)—NH-tert-Butyl 468 H H CH₃ H C(O)—NH-tert-Butyl 469 H H CH₃ H C(O)—NH-tert-Butyl 470 H H CH₃ H C(O)—NH-tert-Butyl 471 H H CH₃ H C(O)—NH-tert-Butyl 472 H H CH₃ H C(O)—NH-tert-Butyl 473 H H CH₃ H C(O)—NH-tert-Butyl 474 H H CH₃ H C(O)—NH-tert-Butyl 475 H H CH₃ H C(O)—NH-tert-Butyl 476 H H CH₃ H C(O)—NH-tert-Butyl 477 H H CH₃ H C(O)—NH-tert-Butyl 478 H H CH₃ H C(O)—NH-tert-Butyl 479 H H CH₃ H C(O)—NH-tert-Butyl 480 H H CH₃ H C(O)—NH-tert-Butyl 481 H H CH₃ H C(O)—NH-tert-Butyl 482 H H CH₃ H C(O)—NH-tert-Butyl 483 H H CH₃ H C(O)—NH-tert-Butyl 484 H H CH₃ H C(O)—NH-tert-Butyl 485 H H CH₃ H C(O)—NH-tert-Butyl 486 H H CH₃ H C(O)—NH-tert-Butyl 487 H H CH₃ H C(O)—NH-tert-Butyl 488 H H CH₃ H C(O)—NH-tert-Butyl 489 H H CH₃ H C(O)—NH-tert-Butyl 490 H H CH₃ H C(O)—NH-tert-Butyl 491 H H CH₃ H C(O)—NH-tert-Butyl 492 H H CH₃ H C(O)—NH-tert-Butyl 493 H H CH₃ H C(O)—NH-tert-Butyl 494 H H CH₃ H C(O)—NH-tert-Butyl 495 H H CH₃ H C(O)—NH-tert-Butyl 496 H H CH₃ H C(O)—NH-tert-Butyl 497 H H CH₃ H C(O)—NH-tert-Butyl 498 H H CH₃ H C(O)—NH-tert-Butyl 499 H H CH₃ H C(O)—NH-tert-Butyl 500 H H CH₃ H C(O)—NH-tert-Butyl 501 H H CH₃ H C(O)—NH-tert-Butyl 502 H H CH₃ H C(O)—NH-tert-Butyl 503 H H CH₃ H C(O)—NH-tert-Butyl 504 H H CH₃ H C(O)—NH-tert-Butyl 505 H H CH₃ H C(O)—NH-tert-Butyl 506 H H CH₃ H C(O)—NH-tert-Butyl 507 H H CH₃ H C(O)—NH-tert-Butyl 508 H H CH₃ H C(O)—NH-tert-Butyl 509 H H CH₃ H C(O)—NH-tert-Butyl 510 H H CH₃ H C(O)—NH-tert-Butyl 511 H H CH₃ H C(O)—NH-tert-Butyl 512 H H CH₃ H C(O)—NH-tert-Butyl 513 H H CH₃ H C(O)—NH-tert-Butyl 514 H H CH₃ H C(O)—NH-tert-Butyl 515 H H CH₃ H C(O)—NH-tert-Butyl 516 H H CH₃ H C(O)—NH-tert-Butyl 517 H H CH₃ H C(O)—NH-tert-Butyl 518 H H CH₃ H C(O)—NH-tert-Butyl 519 H H CH₃ H C(O)—NH-tert-Butyl 520 H H CH₃ H C(O)—NH-tert-Butyl 521 H H CH₃ H C(O)—NH-tert-Butyl 522 H H CH₃ H C(O)—NH-tert-Butyl 523 H H CH₃ H C(O)—NH-tert-Butyl 524 H H CH₃ H C(O)—NH-tert-Butyl 525 H H CH₃ H C(O)—NH-tert-Butyl 526 H H CH₃ H C(O)—NH-tert-Butyl 527 H H CH₃ H C(O)—NH-tert-Butyl 528 H H CH₃ H C(O)—NH-tert-Butyl 529 H H CH₃ H C(O)—NH-tert-Butyl 530 H H CH₃ H C(O)—NH-tert-Butyl 531 H H CH₃ H C(O)—NH-tert-Butyl 532 H H CH₃ H C(O)—NH-tert-Butyl 533 H H CH₃ H C(O)—NH-tert-Butyl 534 H H CH₃ H C(O)—NH-tert-Butyl 535 H H CH₃ H C(O)—NH-tert-Butyl 536 H H CH₃ H C(O)—NH-tert-Butyl 537 H H CH₃ H C(O)—NH-tert-Butyl 538 H H CH₃ H C(O)—NH-tert-Butyl

TABLE 2B Com- Seq. pound ID. No. No. R⁶ R⁷ R⁸ X^(y) X^(z) X¹ X² X³ 390 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 391 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 392 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 393 31 HYP HYP H F L D T 394 31 HYP HYP H F L D T 395 31 HYP HYP H F L D T 396 31 HYP HYP H F L D T 397 31 HYP HYP H F L D T 457 31 HYP HYP H F L D T 458 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 459 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 460 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 461 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 462 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 463 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 464 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 465 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 466 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 467 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 468 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 469 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 470 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 471 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 472 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 473 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 474 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 475 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 476 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 477 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 478 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 479 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 480 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 481 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 482 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 483 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 484 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 485 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 486 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 487 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 488 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 489 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 490 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 491 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 492 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 493 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 494 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 495 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 496 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 497 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 498 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 499 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 500 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 501 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 502 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 503 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 504 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 505 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 506 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 507 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 508 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 509 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 510 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 511 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 512 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 513 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 514 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 515 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 516 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 517 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 518 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 519 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 520 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 521 334 PRO PRO H [3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid] L D T 522 127 PRO PRO H (4-aminomethyl-Phe) dPip L D T 523 127 PRO PRO H (4-aminomethyl-Phe) dPip L D T 524 127 PRO PRO H (4-aminomethyl-Phe) dPip L D T 525 127 PRO PRO H (4-aminomethyl-Phe) dPip L D T 526 127 PRO PRO H (4-aminomethyl-Phe) dPip L D T 527 141 PRO PRO H (3-aminomethyl-Phe) dTic L D T 528 141 PRO PRO H (3-aminomethyl-Phe) dTic L D T 529 141 PRO PRO H (3-aminomethyl-Phe) dTic L D T 530 427 PRO PRO H K MebetaHomoLys L D T 531 427 PRO PRO H K MebetaHomoLys L D T 532 427 PRO PRO H K MebetaHomoLys L D T 533 427 PRO PRO H K MebetaHomoLys L D T 534 427 PRO PRO H K MebetaHomoLys L D T 535 427 PRO PRO H K MebetaHomoLys L D T 536 427 PRO PRO H K MebetaHomoLys L D T 537 427 PRO PRO H K MebetaHomoLys L D T 538 427 PRO PRO H K MebetaHomoLys L D T

TABLE 2C RPMI8866 Ramos ELISA MADCAM FACS Adhesion Adhesion ELISA a4b7 ELISA a4b1 Assay VCAM FACS Compound a4b7 Th mem a4b7/MAdCAM a4b1/VCAM Ratio Assay IC₅₀ Assay IC₅₀ Ratio a4+b7− Th mem No. (nM) IC50 (nM) IC₅₀ (nM) Ramos/RPMI (nM) (nM) b1/b7 (nM) 390 90 13 857 66 8.3 2.0 0.2 391 22 392 Unclear 28 1845 65 18 6.3 0.3 393 635 394 860 395 1521 396 1953 397 2061 457 2163 458 66 42 990 23 6.4 1.5 0.2 459 33 460 107 62 1848 30 14 1.0 0.1 461 93 39 1224 31 18 7.9 0.4 462 68 42 350 8 463 21 464 232 19 1547 80 61 22 0.4 465 132 22 466 164 97 3244 34 14 4.2 0.3 467 43 876 20 8.6 1.0 0.1 468 13 677 51 32 10 0.3 469 5 44 36 10 0.3 470 81 17 1110 66 5.4 1.0 0.2 471 46 29 472 80 473 63 474 74 14 1297 95 19 7.1 0.4 475 74 18 395 22 11 1.1 0.1 476 32 477 124 21 478 34 479 480 92 21 1012 49 16 4.9 0.3 481 67 36 940 26 7.1 1.6 0.2 482 30 26 825 31 4.7 1.5 0.3 4000 483 29 484 45 14 576 41 5.6 1.2 0.2 485 30 486 42 487 65 488 34 489 40 490 41 491 23 492 104 25 493 94 45 1254 28 12 3.5 0.3 494 128 50 161 3 9.4 1.9 0.2 495 30 496 41 497 25 498 517 36 1360 38 12 3.0 0.3 4000 499 41 42 878 21 17 3.1 0.2 500 55 32 1001 31 12 9.9 0.8 501 38 502 Unclear, 148, 1000 102 779 8 503 250 45 504 301 42 505 59 1775 30 19 6.6 0.3 506 289 11004 38 507 189 31 1928 62 18 4.4 0.2 508 46 509 31 510 87 511 182 30 3989 133 11 7.2 0.6 20000 512 64 25 513 31 514 95 515 34 516 98 517 43 10 211 21 7.5 2.1 0.3 607 518 189 18 947 53 36 8.7 0.2 519 27 520 1.2 6 82 14 6.3 1.2 0.2 521 53 10 101 10 9.4 1.5 0.2 522 372 523 434 524 447 525 472 526 611 527 126 43 766 18 4.6 1.1 0.2 528 47 529 47 530 33 15 10557 704 5.3 5.1 1.0 >4000 531 23 532 57 19 7145 370 6.1 6.9 1.1 533 46 17 4235 257 5 6.2 1.2 534 38 23 10839 471 12 10 0.9 >4000 535 77 15 9507 634 8.6 9.7 0.9 536 75 16 7817 479 9.5 8.9 0.9 537 19 6.5 2.5 0.4 538 13 7650 10 13 1.3

TABLE 2X Compound LC-MS Reagent employed to form Linkage Multimer No. (m/z) multimer types(s) type 390 925.7 Diglycolic acid amide homodimer 391 939.0 Pimelic acid amide homodimer 392 973.6 Dodecanedioic acid amide homodimer 393 808.4 Pimelic acid ester homodimer 394 825.4 1,4-Phenylenediacetic acid ester homodimer 395 822.4 Azelaic acid ester homodimer 396 815.4 Suberic acid ester homodimer 397 829.5 Sebacic acid ester homodimer 457 843.5 Dodecanedioic acid ester homodimer 458 938.0 (±)-cis-Cyclopentane-1,2-dicarboxylic acid amide homodimer 459 923.6 1,1-Cyclopropanedicarboxylic acid amide homodimer 460 971.2 1,3-Adamantanedicarboxylic acid amide homodimer 461 961.6 1,3-Dihydroindene-2,2-dicarboxylic acid amide homodimer 462 955.6 1,3-Phenylenediacetic acid amide homodimer 463 967.0 1,4-Naphthalenedicarboxylic acid amide homodimer 464 1004.2  2-Fluoro-5′-methoxybiphenyl-3′,4-dicarboxylic acid amide homodimer 465 980.6 2,2′-Bipyridine-4,4′-dicarboxylic amide homodimer 466 980.6 2,2′-Bipyridine-6,6′-dicarboxylic acid amide homodimer 467 943.2 2,3-Pyrazinedicarboxylic acid amide homodimer 468 937.0 2,4-Dichloro-5-nitro-pyrimidine amine homodimer 469 1017.6  2,5-Diphenylbenzene-1,4-dicarboxylic acid amide homodimer 470 944.8 2,5-Thiophenedicarboxylic acid amide homodimer 471 966.6 2,6-Naphthalenedicarboxylic acid amide homodimer 472 942.6 2,6-Pyridinecarboxylic acid amide homodimer 473 947.8 3,3′-Thiodipropionic acid amide homodimer 474 979.6 4,4-Dibenzoic acid amide homodimer 475 997.8 4,4′-Difluorobiphenyl-2,2′-dicarboxylic acid amide homodimer 476 944.2 4,5-Dicarboxy-1-methyl-1H-imidazole amide homodimer 477 976.2 5-(Trifluoromethyl)benzene-1,3-dicarboxylic acid amide homodimer 478 979.6 Biphenyl-2,3′-dicarboxylic acid amide homodimer 479 979.6 Biphenyl-3,3′-dicarboxylic acid amide homodimer 480 976.6 Biphenyl-3,4′-dicarboxylic acid amide homodimer 481 896.6 Chloroacetyl chloride amide/amine homodimer 482 903.6 Acryloyl chloride amide/amine homodimer 483 910.9 Malonic acid amide homodimer 484 917.9 Succinic acid amide homodimer 485 924.6 Glutaric acid amide homodimer 486 931.6 Adipic acid amide homodimer 487 945.9 Suberic acid amide homodimer 488 952.6 Azelaic acid amide homodimer 489 959.6 Sebacic acid amide homodimer 490 950.0 Chelidamic acid amide homodimer 491 945.0 Cis-1,2-cyclohexanedicarboxylic acid amide homodimer 492 925.6 Aspartic acid amide homodimer 493 925.6 D-Aspartic acid amide homodimer 494 932.2 Glutamic acid amide homodimer 495 932.2 D-Glutamic acid amide homodimer 496 949.0 Homophthalic acid amide homodimer 497 941.6 Isophthalic acid amide homodimer 498 1201.0  Lys(Cbz)-C7-Lys(Cbz) amide homodimer 499 927.8 α,α′-Dibromo-m-xylene amine homodimer 500 927.8 α,α′-Dibromo-p-xylene amine homodimer 501 932.6 Methyliminodiacetic acid amide homodimer 502 1053.2  Pamoic acid amide homodimer 503 1160.0  PEG10-37 atoms amide homodimer 504 1204.0  PEG12-43 atoms amide homodimer 505 1097.6  PEG2-C7-PEG2 amide homodimer 506 1084.6  PEG2-diglycolic acid-PEG2 amide homodimer 507 1038.6  PEG2-diphenic acid-PEG2 amide homodimer 508 969.9 PEG3 linker amide homodimer 509 1027.6  PEG4 linker amide homodimer 510 1072.0  PEG6 linker amide homodimer 511 1137.8  PEG9 linker amide homodimer 512 941.7 Phthalic acid linker amide homodimer 513 945.4 trans-1,2-Cyclohexanedicarboxylic acid amide homodimer 514 937.8 trans-DL-1,2-Cyclopentanedicarboxylic acid amide homodimer 515 963.6 1,3,5-Benzenetricarbonyl trichloride amide homodimer 516 955.6 1,4-Phenylenediacetic acid amide homodimer 517 979.6 diphenic acid amide homodimer 518 914.8 1,3,5-Tris(bromomethyl)benzene amine homotrimer 519 931.0 1,3,5-Cyclohexanetricarboxylic acid (all-cis) amide homotrimer 520 928.6/1392.8 1,3,5-Benzenetricarbonyl trichloride amide homotrimer 521 775.0/969.0/1291.7 (+)-(18-Crown-6)-2,3,11,12-tetracarboxylic acid amide homotramer 522 918.7 Glutaric acid amide homodimer 523 925.6 Adipic acid amide homodimer 524 932.6 Pimelic acid amide homodimer 525 939.6 Suberic acid amide homodimer 526 946.6 Azelaic acid amide homodimer 527 980.6 Pimelic acid amide homodimer 528 987.6 Suberic acid amide homodimer 529 994.6 Azelaic acid amide homodimer 530 950.8 Sebacic acid amide homodimer 531 908.6 Succinic acid amide homodimer 532 929.8 Pimelic acid amide homodimer 533 944.2 Azelaic acid amide homodimer 534 983.4 Diglycolic acid amide homodimer 535 971.2 Diphenic acid amide homodimer 536 1063.2  PEG6 amide homodimer 537 955.2 1,3,5-Benzenetricarbonyl trichloride amide homodimer 538 919.8 1,3,5-Benzenetricarbonyl trichloride amide homotrimer Com- pound Representative structures of Linker moieties Experimental No. (the number 1 represents an attachment point between Linker and monomeric macrocycle) Protocol 390 A, D, Gb, I, M, N, Oa 391 A, D, Gb, I, M, N, Oa 392 A, D, Gb, I, M, N, Oa 393 A, E, Fa, Jb, M, Ob, I, M 394 A, E, Fa, Jb, M, Ob, I, M 395 A, E, Fa, Jb, M, Ob, I, M 396 A, E, Fa, Jb, M, Ob, I, M 397 A, E, Fa, Jb, M, Ob, I, M 457 A, E, Fa, Jb, M, Ob, I, M 458 A, D, Gb, I, M, Nb, Ob 459 A, D, Gb, I, M, Nb, Ob 460 A, D, Gb, I, M, Na, Oa 461 A, D, Gb, I, M, Na, Oa 462 A, D, Gb, I, M, Nc, Ob 463 A, D, Gb, I, M, Na, Oa 464 A, D, Gb, I, M, Na, Oa 465 A, D, Gb, I, M, Na, Oa 466 A, D, Gb, I, M, Nb, Oa 467 A, D, Gb, I, M, Nb, Ob 468

A, D, Gb, I, M, Of 469 A, D, Gb, I, M, Na, Oa 470 A, D, Gb, I, M, Na, Oa 471 A, D, Gb, I, M, Na, Oa 472 A, D, Gb, I, M, Nb, Ob 473 A, D, Gb, I, M, Nc, Ob 474 A, D, Gb, I, M, Na, Oa 475 A, D, Gb, I, M, Na, Oa 476 A, D, Gb, I, M, Nb, Ob 477 A, D, Gb, I, M, Na, Oa 478 A, D, Gb, I, M, Nb, Ob 479 A, D, Gb, I, M, Nb, Ob 480 A, D, Gb, I, M, Nb, Ob 481

A, D, Gb, I, M, Oc 482

A, D, Gb, I, M, Od 483 A, D, Gb, I, M, Nb, Ob 484 A, D, Gb, I, M, Nb, Ob 485 A, D, Gb, I, M, Na, Oa 486 A, D, Gb, I, M, Nb, Ob 487 A, D, Gb, I, M, Na, Oa 488 A, D, Gb, I, M, Na, Oa 489 A, D, Gb, I, M, Na, Oa 490 A, D, Gb, I, M, Na, Oa 491 A, D, Gb, I, M, Nb, Ob 492

A, D, Gb, I, M, Nc, Ob 493

A, D, Gb, I, M, Nc, Ob 494

A, D, Gb, I, M, Nc, Ob 495

A, D, Gb, I, M, Nc, Ob 496 A, D, Gb, I, M, Nb, Ob 497 A, D, Gb, I, M, Nb, Ob 498

A, D, Gb, I, M, Nd, Nc, Ob 499

A, D, Gb, I, M, Oh 500

A, D, Gb, I, M, Oh 501 A, D, Gb, I, M, Nc, Ob 502 A, D, Gb, I, M, Nc, Ob 503

A, D, Gb, I, M, Nb, Ob 504

A, D, Gb, I, M, Nb, Ob 505

A, D, Gb, I, M, Ng, Nb, Ob 506

A, D, Gb, I, M, Ne, Nb, Ob 507

A, D, Gb, I, M, Nf, Nb, Ob 508

A, D, Gb, I, M, Nb, Ob 509

A, D, Gb, I, M, Na, Oa 510

A, D, Gb, I, M, Na, Oa 511

A, D, Gb, I, M, Nc, Ob 512 A, D, Gb, I, M, Nb, Ob 513 A, D, Gb, I, M, Na, Oa 514 A, D, Gb, I, M, Nc, Ob 515

A, D, Gb, I, M, Oa 516 A, D, Gb, I, M, Na, Oa 517 A, D, Gb, I, M, Na, Oa 518 A, D, Gb, I, M, Oh 519 A, D, Gb, I, M, Nb, Ob 520

A, D, Gb, I, M, Oa 521 A, D, Gb, I, M, Nb, Ob 522 A, D, I, M, Og, Ja 523 A, D, I, M, Og, Ja 524 A, D, I, M, Og, Ja 525 A, D, I, M, Og, Ja 526 A, D, I, M, Og, Ja 527 A, D, I, M, Na, Oa 528 A, D, I, M, Na, Oa 529 A, D, I, M, Na, Oa 530 B, A, D, M, Ja, Nc, Ob, I 531 B, A, D, M, Ja, Nb, Ob, I 532 B, A, D, M, Ja, Nc, Ob, I 533 B, A, D, M, Ja, Nc, Ob, I 534 B, A, D, M, Ja, Oa, I 535 B, A, D, M, Ja, Nc, Ob, I 536

B, A, D, M, Ja, Nc, Ob, I 537

B, A, D, M, Ja, Oa, I 538

B, A, D, M, Ja, Nb, Ob, I

TABLE 3 Seq ID Sequence Features 1 Pro Tyr Leu Asp Val 2 Pro His Leu Asp Val 3 Pro Tyr Leu Asp Thr 4 Pro Phe Leu Asp Thr 5 Pro X Leu Asp Thr X is homoPhe 6 Pro X Leu Asp Thr X is beta-cyclohexyl alanine, (S)-2-amino-3-cyclohexylpropionic acid 7 Pro Trp Leu Asp Ile 8 Pro X Leu Asp Thr X is 1-napthylalanine 9 Pro X Leu Asp Thr X is 2-napthylalanine 10 Pro Trp Leu Asp X X is O-benzyl-threonine 11 Pro X Leu Asp Thr X is biphenylalanine 12 Pro X Leu Asp Thr X is O-phenyl-tyrosine 13 Pro X Leu Asp Ile X is 1-napthylalanine 14 Pro X Leu Asp Ile X is 2-napthylalanine 15 Pro X₁ Leu Asp X₂ X₁ is 2-napthylalanine X₂ is O-benzyl-threonine 16 X Trp Leu Asp Thr X is (4S)-fluoro-proline 17 Pro X₁ Leu Asp X₂ X₁ is biphenylalanine X₂ is O-benzyl-threonine 18 Pro X Leu Asp Thr X is O-2-methyl-phenyl-tyrosine 19 Pro X Leu Asp Thr X is O-4-trifluoromethyl-phenyl-tyrosine 20 Pro X Leu Asp Thr X is O-4-methoxy-phenyl-tyrosine 21 Pro X Leu Asp Thr X is O-4-fluoro-phenyl-tyrosine 22 Pro X Leu Asp Thr X is O-2-methoxy-phenyl-tyrosine 23 Pro X Leu Asp Thr X is O-3-methoxy-phenyl-tyrosine 24 Pro X Leu Asp Thr X is O-3-fluoro-phenyl-tyrosine 25 Pro X Leu Asp Thr X is O-3,4-difluoro-phenyl-tyrosine 26 Pro X Leu Asp Thr X is O-3-methyl-phenyl-tyrosine 27 Pro X Leu Asp Thr X is O-3,4-dimethyl-phenyl-tyrosine 28 Pro X Leu Asp Thr X is O-4-methylester-phenyl-tyrosine 29 Pro X Leu Asp Thr X is O-3-methylester-phenyl-tyrosine 30 Pro X Leu Asp Thr X is O-4-carboxylate-phenyl-tyrosine 31 X Phe Leu Asp Thr X is trans-4-hydroxyproline, (2S,4R)-4-hydroxypyrrolidine-2-carboxylic acid 32 Pro X Leu Asp Thr X is metaTyrosine 33 Pro X₁ Leu Asp X₂ X₁ is Ndelta-benzamide-ornithine X₂ is O-benzyl-threonine 34 Pro X₁ Leu Asp X₂ X₁ is Ndelta-acetamide-ornithine X₂ is O-benzyl-threonine 35 Pro X₁ Leu Asp X₂ X₁ is Ndelta-methanesulfonamide-ornithine X₂ is O-benzyl-threonine 36 Pro X₁ Leu Asp X₂ X₁ is Ndelta-ethylcarbamate-ornithine X₂ is O-benzyl-threonine 37 Pro X₁ Leu Asp X₂ X₁ is Ndelta-pentyl amide-ornithine X₂ is O-benzyl-threonine 38 Pro Arg Leu Asp Thr 39 Pro Phe Leu Asp X X is O-methyl-threonine 40 Pro Phe Leu Asp X X is O-ethyl-threonine 41 Pro X Leu Asp Thr X is D-tyrosine 42 Pro X Leu Asp Thr X is (3R)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid 43 X₁ X₂ Leu Asp Thr X₁ is trans-4-hydroxyproline, (2S,4R)-4-hydroxypyrrolidine-2-carboxylic acid X₂ is [3-(3′-pyridyl)-alanine] 44 X Phe Leu Asp Thr X is (4R)-fluoro-proline 45 X₁ X₂ Leu Asp Thr X₁ is (4R)-fluoro-proline X₂ is biphenylalanine 46 X₁ X₂ Leu Asp Thr X₁ is (4R)-fluoro-proline X₂ is [3-(3′-pyridyl)-alanine] 47 X Tyr Leu Asp Thr X is (4R)-fluoro-proline 48 X Tyr Leu Asp Thr X is (4S)-fluoro-proline 49 Pro X Leu Asp Thr X is D-arginine 50 Pro X Leu Asp Thr X is D-pipecolic acid, D-homoPro 51 Pro X Leu Asp Thr X is (3-(4-thiazolyl)-alanine) 52 Pro Tyr Leu Asp Ile 53 Pro X Leu Asp Thr X is 4-aza-phenylalanine 54 Pro Tyr Leu Asp X X is penicillamine, beta, beta-dimethyl-cysteine 55 Pro X Leu Asp Thr X is 2-amino-4-bromo-4-pentenoic acid 56 Pro X Leu Asp Thr X is O-benzyl-trans-4-hydroxyproline 57 Pro X Leu Asp Thr X is Nbeta-Z-2,3-diaminopropionic acid 58 Pro X Leu Asp Thr X is N-tau-benzyl-histidine 59 Pro X Leu Asp Thr X is 4-amino-phenylalanine 60 Pro X Leu Asp Thr X is 4-aza-D-phenylalanine 61 Pro X Leu Asp Thr X is trans-4-hydroxyproline, (2S,4R)-4-hydroxypyrrolidine-2-carboxylic acid 62 Pro X Leu Asp Thr X is D-tryptophan 63 Pro Met Leu Asp Thr 64 Pro X Leu Asp Thr X is D-methionine 65 Pro X Leu Asp Thr X is 4-guanidino-phenylalanine 66 Pro X Leu Asp Thr X is 3-aza-phenylalanine 67 Pro X Leu Asp Thr X is 3-aza-D-phenylalanine 68 Pro X Leu Asp Thr X is norvaline 69 Pro X Leu Asp Thr X is D-norleucine 70 Pro X Leu Asp Thr X is D-lysine 71 Pro X Leu Asp Thr X is D-proline 72 Pro X Leu Asp Thr X is D-ornithine 73 Pro X Leu Asp Thr X is 3-benzothienyl-alanine 74 Pro X Leu Asp Thr X is O-allyl-D-tyrosine 75 Pro X Leu Asp Thr X is O-benzyl-D-serine 76 Pro X Leu Asp Thr X is 3-(4-thiazolyl)-D-alanine 77 Pro X Leu Asp Thr X is 3-benzothienyl-D-alanine 78 Pro X Leu Asp Thr X is 3-(2-thienyl)-D-alanine 79 Pro X Leu Asp Thr X is 4-aminomethyl-phenylalanine 80 Pro X Leu Asp Thr X is Ndelta-dimethyl-D-ornithine 81 Pro X Leu Asp Thr X is 4-amino-D-phenylalanine 82 Pro X Leu Asp Thr X is 4-aminomethyl-D-phenylalanine 83 Pro X Leu Asp Thr X is O-benzyl-D-tyrosine 84 Pro Pro Leu Asp Thr 85 Pro X Leu Asp Thr X is cyclo leucine, 1-aminocyclopentane-1-carboxylic acid 86 Pro X Leu Asp Thr X is aminoindan-2-carboxylic acid 87 Pro X Leu Asp Thr X is O-allyl-tyrosine 88 Pro X Leu Asp Thr X is cyclohexyl glycine 89 Pro Lys Leu Asp Thr 90 Pro X Leu Asp Thr X is 2-aza-D-phenylalanine 91 Pro X Leu Asp Thr X is 2-aza-phenylalanine 92 Pro X Leu Asp Thr X is 2-(2-pyridyl)-4-thiazolyl-alanine 93 Pro X Leu Asp Thr X is 2-(3-pyridyl)-4-thiazolyl-alanine 94 Pro X Leu Asp Thr X is 2-(4-pyridyl)-4-thiazolyl-alanine 95 Pro X Leu Asp Thr X is D-1,2,3,4-tetrahydroisoquinoline-1-carboxylic acid 96 Pro X Leu Asp Thr X is 1-(S)-isoindoline-carboxylic acid 97 Pro Tyr X Leu Asp X is D-threonine Thr 98 Pro Tyr Pro Leu Asp Thr 99 Pro Tyr X Leu Asp X is D-proline Thr 100 Pro Tyr X Leu Asp X is sarcosine, N-methyl glycine Thr 101 Pro Tyr X Leu Asp X is cyclo leucine, 1-Aminocyclopentane-1-carboxylic acid Thr 102 Pro X₁ X₂ Leu Asp X₁ is 3-iodo-phenylalanine Thr X₂ is sarcosine, N-methyl glycine 103 Pro X₁ X₂ Leu Asp X₁ is 4-iodo-phenylalanine Thr X₂ is sarcosine, N-methyl glycine 104 Pro X₁ X₂ Leu Asp X₁ is 3,3-diphenyl-alanine Thr X₂ is sarcosine, N-methyl glycine 105 Pro Phe X Leu Asp X is D-lysine Thr 106 Pro X₁ X₂ Leu Asp X₁ is biphenylalanine Thr X₂ is D-lysine 107 Pro X₁ X₂ Leu Asp X₁ is 3-(4-thiazolyl)-alanine Thr X₂ is D-lysine 108 Pro X₁ X₂ Leu Asp X₁ is 3,3-diphenyl-alanine Thr X₂ is D-lysine 109 Pro Tyr X Leu Asp X is D-lysine Ile 110 Pro Tyr X Leu Asp X is D-arginine Thr 111 Pro Tyr X Leu Asp X is D-serine Thr 112 Pro X₁ X₂ Leu Asp X₁ is biphenylalanine Thr X₂ is sarcosine, N-methyl glycine 113 Pro X₁ X₂ Leu Asp X₁ is 1-napthylalanine Thr X₂ is sarcosine, N-methyl glycine 114 Pro Tyr X Leu Asp X is pipecolic acid, homoPro Thr 115 Pro X₁ X₂ Leu Asp X₁ is 2-iodo-phenylalanine Thr X₂ is sarcosine, N-methyl glycine 116 Pro X₁ X₂ Leu Asp X₁ is 1-napthylalanine Thr X₂ is D-lysine 117 Pro Tyr X₁ Leu Asp X₁ is D-lysine X₂ X₂ is N-methyl threonine 118 Pro Phe X Leu Asp X is sarcosine, N-methyl glycine Thr 119 Pro Tyr X Leu Asp X is (3R)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid Thr 120 Pro Tyr X Leu Asp X is D-pipecolic acid, D-homoPro Thr 121 Pro Phe X Leu Asp X is D-proline Thr 122 Pro X₁ X₂ Leu Asp X₁ is 3,4-dimethoxy-phenylalanine Thr X₂ is D-proline 123 Pro X₁ X₂ Leu Asp X₁ is 3,4,5-trifluoro-phenylalanine Thr X₂ is D-proline 124 Pro X₁ X₂ Leu Asp X₁ is 3,5-dibromo-tyrosine Thr X₂ is D-proline 125 Pro Phe X Leu Asp X is D-pipecolic acid, D-homoPro Thr 126 Pro X₁ X₂ Leu Asp X₁ is 3-(4-thiazolyl)-alanine Thr X₂ is D-pipecolic acid, D-homoPro 127 Pro X₁ X₂ Leu Asp X₁ is 4-aminomethyl-phenylalanine Thr X₂ is D-pipecolic acid, D-homoPro 128 Pro X₁ X₂ Leu Asp X₁ is 2-iodo-phenylalanine Thr X₂ is D-pipecolic acid, D-homoPro 129 Pro X₁ X₂ Leu Asp X₁ is 2-phenyl-phenylalanine Thr X₂ is D-pipecolic acid, D-homoPro 130 Pro X₁ X₂ Leu Asp X₁ is 2-(2-methoxy-phenyl)-phenylalanine Thr X₂ is D-pipecolic acid, D-homoPro 131 Pro X₁ X₂ Leu Asp X₁ is 2-(3-methoxy-phenyl)-phenylalanine Thr X₂ is D-pipecolic acid, D-homoPro 132 Pro X₁ X₂ Leu Asp X₁ is 2-(4-methoxy-phenyl)-phenylalanine Thr X₂ is D-pipecolic acid, D-homoPro 133 Pro X₁ X₂ Leu Asp X₁ is biphenylalanine Thr X₂ is D-pipecolic acid, D-homoPro 134 Pro Tyr X Leu Asp X is trans-4-hydroxyproline, (2S,4R)-4-hydroxypyrrolidine-2-carboxylic Thr acid 135 Pro Tyr X Leu Asp X is trans-D-4-hydroxyproline, (2R,4S)-4-hydroxypyrrolidine-2- Thr carboxylic acid 136 Pro Tyr X Leu Asp X is cis-D-4-Hydroxyproline, (2R,4R)-4-Hydroxypyrrolidine-2-carboxylic Thr acid 137 X₁ X₂ X₃ Leu Asp Thr X₁ is D-proline X₂ is D-tyrosine X₃ is D-pipecolic acid, D-homoPro 138 Pro X₁ X₂ Leu Asp X₁ is 1-napthylalanine Thr X₂ is D-pipecolic acid, D-homoPro 139 Pro X₁ X₂ Leu Asp X₁ is 2-napthylalanine Thr X₂ is D-pipecolic acid, D-homoPro 140 Pro X₁ X₂ Leu Asp X₁ is 4-aminomethyl-phenylalanine Thr X₂ is (3R)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid 141 Pro X₁ X₂ Leu Asp X₁ is 3-aminomethyl-phenylalanine Thr X₂ is (3R)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid 142 Pro X₁ X₂ Leu Asp X₁ is 3-aminomethyl-D-phenylalanine Thr X₂ is (3R)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid 143 Pro X₁ X₂ Leu Asp X₁ is N-methyl tyrosine Thr X₂ is D-pipecolic acid, D-homoPro 144 Pro Tyr X₁ Leu Asp X₁ is D-pipecolic acid, D-homoPro X₂ X₂ is allo-threonine, (2S,3S)-2-amino-3-hydroxybutyric acid 145 Pro Tyr X₁ X₂ Asp X₁ is D-pipecolic acid, D-homoPro Thr X₂ is beta-tert-butyl alanine, neopentylglycine 146 Pro X₁ X₂ Leu Asp X₁ is 3-(4-thiazolyl)-alanine Thr X₂ is trans-D-4-hydroxyproline, (2R,4S)-4-hydroxypyrrolidine-2- carboxylic acid 147 Pro X₁ X₂ Leu Asp X₁ is 4-aminomethyl-phenylalanine Thr X₂ is trans-D-4-hydroxyproline, (2R,4S)-4-hydroxypyrrolidine-2- carboxylic acid 148 Pro Tyr X Leu Asp X is D-pipecolic acid, D-homoPro Ile 149 Pro Tyr X Leu Asp X is N-methyl-D-lysine Ile 150 Pro Tyr X Leu Asp X is D-norleucine Thr 151 Pro Phe X Leu Asp X is trans-D-4-hydroxyproline, (2R,4S)-4-hydroxypyrrolidine-2- Thr carboxylic acid 152 Pro Tyr X Leu Asp X is N-methyl-D-arginine Thr 153 Pro Tyr Gly Leu Asp Thr 154 Pro Tyr Ala Leu Asp Thr 155 Pro Tyr X Leu Asp X is D-alanine Thr 156 Pro Met Gly Leu Asp Thr 157 Pro X₁ X₂ Leu Asp X₁ is O-allyl-tyrosine Thr X₂ is sarcosine, N-methyl glycine 158 Pro X Gly Leu Asp X is O-allyl-tyrosine Thr 159 Pro X₁ X₂ Leu Asp X₁ is 3-(4-thiazolyl)-alanine Thr X₂ is sarcosine, N-methyl glycine 160 Pro X Gly Leu Asp X is 4-aminomethyl-phenylalanine Thr 161 Pro X₁ X₂ Leu Asp X₁ is O-allyl-tyrosine Thr X₂ is D-valine 162 Pro X₁ X₂ Leu Asp X₁ is O-allyl-tyrosine Thr X₂ is D-serine 163 Pro X₁ X₂ Leu Asp X₁ is O-allyl-tyrosine Thr X₂ is D-alanine 164 Pro X Pro Leu Asp X is O-allyl-tyrosine Thr 165 Pro X₁ X₂ Leu Asp X₁ is O-allyl-tyrosine Thr X₂ is D-proline 166 Pro X₁ X₂ Leu Asp X₁ is 3-(4-thiazolyl)-alanine Thr X₂ is D-valine 167 Pro X₁ X₂ Leu Asp X₁ is 3-(4-thiazolyl)-alanine Thr X₂ is D-serine 168 Pro X₁ X₂ Leu Asp X₁ is 3-(4-thiazolyl)-alanine Thr X₂ is D-alanine 169 Pro X Pro Leu Asp X is 3-(4-thiazolyl)-alanine Thr 170 Pro X₁ X₂ Leu Asp X₁ is 3-(4-thiazolyl)-alanine Thr X₂ is D-proline 171 Pro X Pro Leu Asp X is 4-aminomethyl-phenylalanine Thr 172 Pro X₁ X₂ Leu Asp X₁ is 4-aminomethyl-phenylalanine Thr X₂ is D-proline 173 Pro X Pro Leu Asp X is cyclo leucine, 1-aminocyclopentane-1-carboxylic acid Thr 174 Pro X₁ X₂ Leu Asp X₁ is 2-(2-pyridyl)-4-thiazolyl-alanine Thr X₂ is sarcosine, N-methyl glycine 175 Pro X₁ X₂ Leu Asp X₁ is 2-(2-pyridyl)-4-thiazolyl-alanine Thr X₂ is D-proline 176 Pro X₁ X₂ Leu Asp X₁ is 2-(3-pyridyl)-4-thiazolyl-alanine Thr X₂ is sarcosine, N-methyl glycine 177 Pro X₁ X₂ Leu Asp X₁ is 2-(3-pyridyl)-4-thiazolyl-alanine Thr X₂ is D-proline 178 Pro X₁ X₂ Leu Asp X₁ is 2-(4-pyridyl)-4-thiazolyl-alanine Thr X₂ is D-proline 179 Pro X₁ X₂ Leu Asp X₁ is 3-(2-aminobenzyl-4-thiazolyl)-alanine Thr X₂ is sarcosine, N-methyl glycine 180 Pro X₁ X₂ Leu Asp X₁ is 2-(amino-benzyl)-4-thiazolyl-alanine Thr X₂ is D-proline 181 Pro X₁ X₂ Leu Asp Ile X₁ is D-tyrosine X₂ is D-pipecolic acid, D-homoPro 182 Pro X₁ X₂ Leu Asp X₁ is 2-aminomethyl-phenylalanine Thr X₂ is azetidine-2-carboxylic acid 183 Pro Tyr X₁ Leu Asp X₁ is D-pipecolic acid, D-homoPro X₂ X₂ is 2-aminobutyric acid 184 Pro X₁ X₂ Leu Asp X₃ X₁ is 3-aminomethyl-phenylalanine X₂ is (3R)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid X₃ is 2-aminobutyric acid 185 Pro X₁ X₂ Leu Asp X₁ is 2,4-dichloro-phenylalanine Thr X₂ is D-pipecolic acid, D-homoPro 186 Pro X₁ X₂ Leu Asp X₁ is 3-phenyl-D-phenylalanine Thr X₂ is D-pipecolic acid, D-homoPro 187 Pro X₁ X₂ Leu Asp X₁ is 3-(5-quinolinyl)-D-phenylalanine Thr X₂ is D-pipecolic acid, D-homoPro 188 Pro Tyr X Leu Asp X is beta-homolysine Thr 189 Pro Tyr X Leu Asp X is beta-homoproline Thr 190 Pro Tyr X Leu Asp X is beta-homolysine Thr 191 Pro Tyr X Leu Asp X is anthranilic acid, 2-aminobenzoic acid Thr 192 Pro Phe X Leu Asp X is beta-homolysine Thr 193 Pro X₁ X₂ Leu Asp X₁ is 3-(4-thiazolyl)-alanine Thr X₂ is beta-homolysine 194 Pro X₁ X₂ Leu Asp X₁ is 4-aminomethyl-phenylalanine Thr X₂ is beta-homolysine 195 Pro Tyr X₁ Leu Asp X₁is beta-homolysine X₂ X₂ is O-benzyl-threonine 196 Pro X₁ X₂ Leu Asp X₁ is N-methyl tyrosine Thr X₂ is D-beta-homolysine 197 Pro X₁ X₂ Leu Asp X₁ is 1-napthylalanine Thr X₂ is beta-homolysine 198 Pro X₁ X₂ Leu Asp X₁ is 2-napthylalanine Thr X₂ is beta-homolysine 199 Pro X₁ X₂ Leu Asp X₁ is biphenylalanine Thr X₂ is beta-homolysine 200 Pro X₁ X₂ Leu Asp X₁ is 2-iodo-phenylalanine Thr X₂ is beta-homolysine 201 Pro X₁ X₂ Leu Asp X₁ is 2-(2,5-dimethyl-isoxazole)-phenylalanine Thr X₂ is beta-homolysine 202 Pro X₁ X₂ Leu Asp X₁ is 2-phenyl-phenylalanine Thr X₂ is beta-homolysine 203 Pro X₁ X₂ Leu Asp X₁ is (2-piperazinyl-2-phenyl)-phenylalanine Thr X₂ is beta-homolysine 204 Pro X₁ X₂ Leu Asp X₁ is beta-cyclohexyl alanine, (S)-2-amino-3-cyclohexylpropionic acid Thr X₂ is beta-homolysine 205 Pro Trp X Leu Asp X is beta-homolysine Thr 206 Pro X₁ X₂ Leu Asp X₁ is D-tryptophan Thr X₂ is beta-homolysine 207 Pro X₁ X₂ Leu Asp X₁ is 3-aminomethyl-phenylalanine Thr X₂ is beta-homolysine 208 Pro X₁ X₂ Leu Asp X₁ is 4-aminomethyl-D-phenylalanine Thr X₂ is beta-homolysine 209 Pro X₁ X₂ Leu Asp Ile X₁ is 4-aminomethyl-phenylalanine X₂ is beta-homolysine 210 Pro Tyr X Leu Asp X is beta-D-homolysine Ile 211 Pro X₁ X₂ Leu Asp X₁ is D-arginine Thr X₂ is beta-homolysine 212 Pro X₁ X₂ Leu Asp X₁ is 4-aminomethyl-phenylalanine-reduced Thr X₂ is beta-homolysine 213 Pro X₁ X₂ Leu Asp Ile X₁ is 3-(4-thiazolyl)-alanine X₂ is beta-D-homolysine 214 Pro Phe X Leu Asp X is beta-D-homolysine Ile 215 Pro X₁ X₂ Leu Asp X₁ is 3-(4-thiazolyl)-alanine Thr X₂ is N-methyl beta-homolysine 216 Pro X₁ X₂ Leu Asp X₁ is 4-aminomethyl-phenylalanine Thr X₂ is N-methyl beta-homolysine 217 Pro X₁ X₂ Leu Asp Ile X₁ is 3-(4-thiazolyl)-alanine X₂ is beta-homolysine 218 Pro X₁ X₂ Leu Asp X₁ is (3S)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid Thr X₂ is beta-homolysine 219 Pro X₁ X₂ Leu Asp X₁ is (3R)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid Thr X₂ is beta-homolysine 220 Pro X₁ X₂ Leu Asp X₁ is (3R)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid Thr X₂ is beta-D-homolysine 221 Pro Tyr X Leu Asp X is beta-homoisoleucine Thr 222 Pro X₁ X₂ Leu Asp X₁ is 4-aminomethyl-phenylalanine Thr X₂ is beta-homoproline 223 Pro Tyr X Leu Asp X is beta-D-homoproline Thr 224 Pro X₁ X₂ Leu Asp X₁ is 4-aminomethyl-phenylalanine Thr X₂ is beta-D-homoproline 225 Pro Arg X Leu Asp X is beta-homolysine Thr 226 Pro Phe X Leu Asp X is N-methyl beta-homolysine Thr 227 Pro X₁ X₂ Leu Asp X₁ is phenlyalanine-reduced Thr X₂ is beta-homolysine 228 Pro X₁ X₂ Leu Asp X₁ is 3-aminomethyl-D-phenylalanine Thr X₂ is beta-homolysine 229 Pro X₁ X₂ Leu Asp X₁ is {2-[3-(1-piperazinyl)phenyl]-phenylalanine}-beta-homolysine Thr X₂ is beta-homolysine 230 Pro X₁ X₂ Leu Asp X₁ is 3-(4-thiazolyl)-D-alanine Thr X₂ is beta-homolysine 231 Pro X₁ X₂ Leu Asp X₁ is 2-bromo-phenylalanine Thr X₂ is beta-homolysine 232 Pro X₁ X₂ Leu Asp X₁ is 2-chloro-phenylalanine Thr X₂ is beta-homolysine 233 Pro X₁ X₂ Leu Asp X₁ is 2-fluoro-phenylalanine Thr X₂ is beta-homolysine 234 Pro X₁ X₂ Leu Asp X₁ is 2-trifluoromethyl-phenlyalanine Thr X₂ is beta-homolysine 235 Pro X₁ X₂ Leu Asp X₁ is 2,4-dichloro-phenlyalanine Thr X₂ is beta-homolysine 236 Pro X₁ X₂ Leu Asp X₁ is 2-aminomethyl-phenylalanine Thr X₂ is beta-homolysine 237 Pro X₁ X₂ Leu Asp X₁ is 2-(4-quinolinyl)-phenylalanine Thr X₂ is beta-homolysine 238 Pro X₁ X₂ Leu Asp X₁ is 2-(5-quinolinyl)-phenylalanine Thr X₂ is beta-homolysine 239 Pro X₁ X₂ Leu Asp X₁ is 2-(3-quinolinyl)-phenylalanine Thr X₂ is beta-homolysine 240 Pro X₁ X₂ Leu Asp X₁ is D-homophenylalanine Thr X₂ is beta-homolysine 241 Pro X₁ X₂ Leu Asp X₁ is 2-iodo-D-phenylalanine Thr X₂ is beta-homolysine 242 Pro X₁ X₂ Leu Asp X₁ is 2-phenyl-D-phenylalanine Thr X₂ is beta-homolysine 243 Pro X₁ X₂ Leu Asp X₁ is (2-piperazinyl-2-phenyl)-D-phenylalanine Thr X₂ is beta-homolysine 244 Pro Tyr X Leu Asp X is beta-homolysine Ile 245 Pro Tyr X Leu Asp X is beta-homolysine Val 246 Pro X₁ X₂ Leu Asp Ile X₁ is D-tyrosine X₂ is beta-homolysine 247 Pro X₁ X₂ Leu Asp Ile X₁ is 4-aminomethyl-D-phenylalanine X₂ is beta-homolysine 248 Pro X₁ X₂ Leu Asp X₁ is 4-aminomethyl-phenylalanine Val X₂ is beta-homolysine 249 Pro X₁ X₂ Leu Asp X₁ is 3-iodo-phenylalanine Thr X₂ is beta-homolysine 250 Pro X₁ X₂ Leu Asp X₁ is 3-phenyl-phenylalanine Thr X₂ is beta-homolysine 251 Pro X₁ X₂ Leu Asp X₁ is 3-(2-methoxy-phenyl)-phenylalanine Thr X₂ is beta-homolysine 252 Pro X₁ X₂ Leu Asp X₁ is 3-(2,6-dimthoxy-phenyl)-phenylalanine Thr X₂ is beta-homolysine 253 Pro X₁ X₂ Leu Asp X₁ is 3-(2-trifluoromethoxy-phenyl)-phenylalanine Thr X₂ is beta-homolysine 254 Pro X₁ X₂ Leu Asp X₁ is 4-iodo-phenylalanine Thr X₂ is beta-homolysine 255 Pro X₁ X₂ Leu Asp X₁ is 4-(2-methoxy-phenyl)-phenylalanine Thr X₂ is beta-homolysine 256 Pro X₁ X₂ Leu Asp X₁ is 4-(2-trifluoromethoxy-phenyl)-phenylalanine Thr X₂ is beta-homolysine 257 Pro X₁ X₂ Leu Asp X₁ is alpha-methyl-phenylalanine, (S)-(—)-2-amino-2-methyl-3- Thr phenylpropionic acid X₂ is beta-homolysine 258 Pro X₁ X₂ Leu Asp X₁ is N-methyl phenylalanine Thr X₂ is beta-homolysine 259 Pro X₁ X₂ Leu Asp X₁ is 3-(2,6-dimethyl-phenyl)-phenylalanine Thr X₂ is beta-homolysine 260 Pro X₁ X₂ Leu Asp X₁ is 3-(quinolin-4-yl)-phenylalanine Thr X₂ is beta-homolysine 261 Pro X₁ X₂ Leu Asp X₁ is 3-(3,4-difluoro-phenyl)-phenylalanine Thr X₂ is beta-homolysine 262 Pro X₁ X₂ Leu Asp X₁ is 4-(2,6-dimethyl-phenyl)-phenylalanine Thr X₂ is beta-homolysine 263 Pro X₁ X₂ Leu Asp X₁ is 4-(2-chloro-6-methoxy-phenyl)-phenylalanine Thr X₂ is beta-homolysine 264 Pro X₁ X₂ Leu Asp X₁ is 3-(4-thiazolyl)-alanine Thr X₂ is beta-homolysine 265 Pro X₁ X₂ Leu Asp X₁ is 2-(4-[1-piperazinyl)phenyl)-phenylalanine Thr X₂ is beta-homolysine 266 Pro X₁ X₂ Leu Asp X₁ is 2-(2,6-dimethylphenyl)-phenylalanine Thr X₂ is beta-homolysine 267 Pro X₁ X₂ Leu Asp X₁ is 2-(benzolthiazol-5-yl)-phenylalanine Thr X₂ is beta-homolysine 268 Pro X₁ X₂ Leu Asp X₁ is homophenylalanine Thr X₂ is beta-homolysine 269 Pro X₁ X₂ Leu Asp X₁ is piperidine-4-amino-4-carboxylic acid Thr X₂ is beta-homolysine 270 Pro X₁ X₂ Leu Asp X₁ is 2-(2,5-dimethyl-isoxazole)-D-phenylalanine Thr X₂ is beta-homolysine 271 Pro X₁ X₂ Leu Asp X₁ is D-tyrosine Val X₂ is beta-homolysine 272 Pro X₁ X₂ Leu Asp X₁ is 4-aminomethyl-D-phenylalanine Thr X₂ is beta-homolysine 273 Pro X₁ X₂ Leu Asp X₁ is 2-(2-chloro-6-methoxyphenyl)-phenylalanine Thr X₂ is beta-homolysine 274 Pro X₁ X₂ Leu Asp X₁ is 2-indanylglycine Thr X₂ is beta-homolysine 275 Pro X₁ X₂ Leu Asp X₁ is 2-indanyl-D-glycine Thr X₂ is beta-homolysine 276 Pro X₁ X₂ Leu Asp X₁ is 2-aminotetraline-2-carboxylic acid Thr X₂ is beta-homolysine 277 Pro Tyr X₁ Leu Asp X₁ is beta-homolysine X₂ X₂ is allo-isoleucine, (2S,3R)-2-amino-3-methylpentanoic acid 278 Pro X₁ X₂ Leu Asp X₃ X₁ is D-tyrosine X₂ is beta-homolysine X₃ is allo-isoleucine, (2S,3R)-2-amino-3-methylpentanoic acid 279 Pro X₁ X₂ Leu Asp X₃ X₁ is 4-aminomethyl-phenylalanine X₂ is beta-homolysine X₃ is allo-isoleucine, (2S,3R)-2-amino-3-methylpentanoic acid 280 Pro X₁ X₂ Leu Asp X₁ is 2-(2,5-bis(trifluoromethyl)phenyl)-phenylalanine Thr X₂ is beta-homolysine 281 Pro X₁ X₂ Leu Asp X₁ is 2-(2,5-bis(trifluoromethyl)phenyl)-phenylalanine Thr X₂ is beta-homolysine 282 Pro X₁ X₂ Leu Asp X₁ is aminoindan-2-carboxylic acid Thr X₂ is beta-homolysine 283 Pro Pro X Leu Asp X is beta-homolysine Thr 284 Pro X₁ X₂ Leu Asp X₁ is D-proline Thr X₂ is beta-homolysine 285 Pro X₁ X₂ Leu Asp X₁ is pipecolic acid, homoPro Thr X₂ is beta-homolysine 286 Pro X₁ X₂ Leu Asp X₁ is 2-(3-pyridyl)-phenylalanine Thr X₂ is beta-homolysine 287 Pro X₁ X₂ Leu Asp X₁ is 2-(4-pyridyl)-phenylalanine Thr X₂ is beta-homolysine 288 Pro X₁ X₂ Leu Asp X₁ is 2-(3-bromo-2-pyridyl)-phenylalanine Thr X₂ is beta-homolysine 289 Pro Tyr X Leu Asp X is beta-D-homolysine Thr 290 Pro X₁ X₂ Leu Asp X₁ is N-benzyl-glycine Thr X₂ is beta-homolysine 291 Pro X₁ X₂ Leu Asp X₁ is 2-(2-bromo-3-pyridyl)-phenylalanine Thr X₂ is beta-homolysine 292 Pro X₁ X₂ Leu Asp X₁ is 3-(2-chloro-6-methoxy-phenyl)-phenylalanine Thr X₂ is beta-homolysine 293 Pro X₁ X₂ Leu Asp X₁ is 3-(benzothiazol-5-yl)-phenylalanine Thr X₂ is beta-homolysine 294 Pro X₁ X₂ Leu Asp X₁ is 2-aminomethyl-phenylalanine Thr X₂ is N-methyl beta-homolysine 295 Pro X₁ X₂ Leu Asp X₁ is 2-aminomethyl-D-phenylalanine Thr X₂ is N-methyl beta-homolysine 296 Pro X₁ X₂ Leu Asp X₁ is 3-(4-thiazolyl)-D-alanine Thr X₂ is N-methyl beta-homolysine 297 Pro X₁ X₂ Leu Asp X₁ is 2-(2-trifluoromethoxy-phenyl)-D-phenylalanine Thr X₂ is N-methyl beta-homolysine 298 Pro X₁ X₂ Leu Asp X₁ is (3S)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid Thr X₂ is N-methyl beta-homolysine 299 Pro X₁ X₂ Leu Asp X₁ is (3R)-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid Thr X₂ is N-methyl beta-homolysine 300 Pro X₁ X₂ Leu Asp X₁ is 2-(5-quinolinyl)-D-phenylalanine Thr X₂ is beta-homolysine 301 Pro Tyr X₁ Leu Asp X₁ is beta-homolysine X₂ X₂ is allo-threonine, (2S,3S)-2-amino-3-hydroxybutyric acid 302 Pro Tyr X₁ Leu Asp X₁ is N-methyl beta-homolysine X₂ X₂ is allo-threonine, (2S,3S)-2-amino-3-hydroxybutyric acid 303 Pro X₁ X₂ Leu Asp X₁ is N-methyl tyrosine Thr X₂ is N-methyl beta-homolysine 304 Pro X₁ X₂ Leu Asp X₃ X₁ is N-methyl tyrosine X₂ is N-methyl beta-homolysine X₃ is allo-threonine, (2S,3S)-2-amino-3-hydroxybutyric acid 305 Pro X₁ X₂ Leu Asp X₁ is N-methyl phenylalanine Thr X₂ is N-methyl beta-homolysine 306 Pro X₁ X₂ Leu Asp X₁ is 2-fluoro-phenylalanine Thr X₂ is N-methyl beta-homolysine 307 Pro X₁ X₂ Leu Asp X₁ is 2-fluoro-N-methyl phenylalanine Thr X₂ is N-methyl beta-homolysine 308 Pro X₁ X₂ Leu Asp X₁ is 2,4-dichloro-phenylalanine Thr X₂ is N-methyl beta-homolysine 309 Pro X₁X₂ Leu Asp X₁ is 2,4-dichloro-N-methyl phenylalanine Thr X₂ is N-methyl beta-homolysine 310 Pro X₁ X₂ Leu Asp X₁ is 2-aminomethyl-N-methyl-phenylalanine Thr X₂ is N-methyl beta-homolysine 311 Pro X₁ X₂ Leu Asp X₁ is 3-(2,6-dimethoxy-phenyl)-D-phenylalanine Thr X₂ is beta-homolysine 312 Pro X₁ X₂ Leu Asp X₁ is 3-(4-quinolinyl)-D-phenylalanine Thr X₂ is beta-homolysine 313 Pro X₁ X₂ Leu Asp X₁ is beta-homolysine Thr X₂ is azetidine-2-carboxylic acid 314 Pro X₁ X₂ Leu Asp X₁ is 3-phenyl-D-phenylalanine Thr X₂ is beta-homolysine 315 Pro X₁ X₂ Leu Asp X₁ is 3-(2-trifluoromethoxy-phenyl)-D-phenylalanine Thr X₂ is beta-homolysine 316 Pro X₁ X₂ Leu Asp X₁ is 3-(2-methoxy-phenyl)-D-phenylalanine Thr X₂ is beta-homolysine 317 Pro X₁ X₂ Leu Asp X₁ is 2-(5-quinolinyl)-N-methyl-phenylalanine Thr X₂ is N-methyl beta-homolysine 318 Pro Phe X Leu Asp X is beta-homonorleucine Thr 319 Pro Phe X Leu Asp X is N-alpha-methyl-N-epsilon-dimethyl-beta-homolysine Thr 320 Pro X₁ X₂ Leu Asp X₁ is N-methyl phenylalanine Thr X₂ is N-alpha-methyl-N-epsilon-dimethyl-beta-homolysine 321 Pro Met X Leu Asp X is N-methyl beta-homolysine Thr 322 Pro X₁ X₂ Leu Asp X₁ is 2-indanylglycine Thr X₂ is N-methyl beta-homolysine 323 Pro X₁ X₂ Leu Asp X₁ is homophenylalanine Thr X₂ is N-methyl beta-homolysine 324 Pro X₁ X₂ Leu Asp X₁ is O-benzyl-trans-4-hydroxyproline Thr X₂ is N-methyl beta-homolysine 325 Pro X₁ X₂ Leu Asp X₁ is cis-2-aminocyclohexanecarboxylic acid Thr X₂ is N-methyl beta-homolysine 326 Pro X₁ X₂ Leu Asp X₁ is N-methyl methionine Thr X₂ is N-methyl beta-homolysine 327 Pro X₁ X₂ Leu Asp X₁ is beta-homolysine Thr X₂ is beta-homolysine 328 Pro X₁ X₂ Leu Asp X₁ is beta-homophenylalanine Thr X₂ is N-methyl beta-homolysine 329 Pro X₁ X₂ Leu Asp X₁ is beta-homomethionine Thr X₂ is N-methyl beta-homolysine 330 Pro Tyr X Leu Asp X is 3-aminomethyl-4-bromo-benzoic acid Thr 331 Pro Tyr X Leu Asp X is 3-aminomethyl-4-(4-aza-phenyl)-benzoic acid Thr 332 Pro Tyr X Leu Asp X is 3-aminomethyl-4-(2,5-dimethyl-isoxazole)-benzoic acid Thr 333 Pro Tyr X Leu Asp X is 3-aminomethyl-4-(3-aminomethyl-phenyl)-benzoic acid Thr 334 Pro X Leu Asp Thr X is 3-aminomethyl-4-(4-(1-piperazinyl)-phenyl)-benzoic acid 335 Pro X Leu Asp Thr X is 3-aminomethyl-4-(4-quinolinyl)-benzoic acid 336 Pro X Leu Asp Thr X is 3-aminomethyl-4-bromo-benzoic acid 337 Pro X Leu Asp Thr X is 3-aminomethyl-4-(2,5-dimethyl-isoxazole)-benzoic acid 338 X₁ X₂ Leu Asp Thr X₁ is D-proline X₂ is 3-aminomethyl-4-(4-pyridyl)-benzoic acid 339 Pro X Leu Asp Thr X is 3-aminomethyl-(4-methylpyrazole-3-yl)-benzoic acid 340 Pro X Leu Asp Thr X is 3-aminomethyl-4-(3-quinolinyl)-benzoic acid 341 Pro X Leu Asp Thr X is 3-aminomethyl-4-(5-quinolinyl)-benzoic acid 342 Pro X Leu Asp Thr X is 3-aminomethyl-4-(2-(1-piperazinyl)phenyl)-benzoic acid 343 Pro X Leu Asp Thr X is 3-aminomethyl-4-(3-(1-piperzainyl)phenyl)-benzoic acid 344 Pro X Leu Asp Thr X is 3-aminomethyl-4-(2-(3-(piperidin-4-ylmethoxy)phenyl))-benzoic acid 345 Pro X Leu Asp Thr X is 3-aminomethyl-4-(4-pyridyl)-benzoic acid 346 Pro X₁ Leu Asp X₂ X₁ is 3-aminomethyl-4-(4-pyridyl)-benzoic acid X₂ is O-benzyl-threonine 347 Pro X₁ Leu Asp X₂ X₁ is 3-aminomethyl-4-(4-quinolinyl)-benzoic acid X₂ is allo-threonine, (2S,3S)-2-amino-3-hydroxybutyric acid 348 Pro X Leu Asp Thr X is 3-aminomethyl-4-(4-(1-piperazinyl)phenyl))-benzoic acid 349 Pro X₁ X₂ Asp Thr X₁ is 3-aminomethyl-4-(4-quinolinyl))-benzoic acid X₂ is beta-tert-butyl alanine, neopentylglycine 350 Pro X Leu Asp Thr X is N-benzyl-3-aminomethyl-benzoic acid 351 Pro X Leu Asp Thr X is 3-aminomethyl-benzoic acid 352 Pro X Leu Asp Thr X is 3-aminomethyl-5-bromo-benzoic acid 353 Pro X Leu Asp Thr X is 3-aminomethyl-6-bromo-benzoic acid 354 Pro X Leu Asp Thr X is 3-aminomethyl-5-(4-aza-phenyl)-benzoic acid 355 Pro X Leu Asp Thr X is 3-aminomethyl-4-(3-thiophenyl)-benzoic acid 356 Pro X Leu Asp Thr X is 3-aminomethyl-4-(4-N,N-dimethyl-carboxamide-phenyl)-benzoic acid 357 Pro X Leu Asp Thr X is 3-aminomethyl-4-(4-aza-phenyl)-benzoic acid 358 Pro X Leu Asp Thr X is 3-aminomethyl-4-(3-aza-phenyl)-benzoic acid 359 Pro X Leu Asp Thr X is 3-aminomethyl-4-(4-hydroxy-phenyl)-benzoic acid 360 Pro X Leu Asp Thr X is 3-aminomethyl-4-(5-(2,4-dimethyl)thiazole)-benzoic acid 361 Pro X Leu Asp Thr X is 3-aminomethyl-4-(3-N,N-dimethylaniline)-benzoic acid 362 Pro X Leu Asp Thr X is 3-aminomethyl-4-(2-fluoro-pyridyl)-benzoic acid 363 Pro X Leu Asp Thr X is 3-aminomethyl-4-(5-pyrimidinyl)-benzoic acid 364 Pro X Leu Asp Thr X is 3-aminomethyl-4-(3-N,N-dimethyl-diarylether)-benzoic acid 365 Pro X Leu Asp Thr X is 3-aminomethyl-4-(3-trifluoromethyl-phenyl)-benzoic acid 366 Pro X Leu Asp Thr X is 3-aminomethyl-4-(2,5-dimethoxy-phenyl)-benzoic acid 367 Pro X Leu Asp Thr X is 3-aminomethyl-4-((2,3,4-tri-methoxy)-phenyl)-benzoic acid 368 Pro X Leu Asp Thr X is 3-aminomethyl-4-(4-carboxy)-phenyl-benzoic acid 369 Pro X Leu Asp Thr X is 3-aminomethyl-4-piperonyl-benzoic acid 370 Pro X Leu Asp Thr X is 3-aminomethyl-4-piperidinyl-benzoic acid 371 Pro X Leu Asp Thr X is 3-aminomethyl-4-morpholinyl-benzoic acid 372 Pro X Leu Asp Thr X is 3-aminomethyl-4-(N,N-dimethyl)-benzoic acid 373 Pro X Leu Asp Thr X is 3-aminomethyl-4-(2-aminomethylphenyl)-benzoic acid 374 Pro X Leu Asp Thr X is 3-aminomethyl-4-(3-aminomethylphenyl)-benzoic acid 375 Pro X Leu Asp Thr X is 3-aminomethyl-4-(4-aminomethylphenyl)-benzoic acid 376 Pro X₁ Leu Asp X₂ X₁ is 3-aminomethyl-4-(4-quinolinyl)-benzoic acid X₂ is 2-aminobutyric acid 377 X₁ X₂ Leu Asp Thr X₁ is norvaline X₂ is 3-aminomethyl-4-(4-quinolinyl)-benzoic acid 378 Pro X Leu Asp Thr X is N-methyl-3-aminomethyl-benzoic acid 379 Pro X Leu Asp Thr X is N-methyl-3-aminomethyl-4-(4-quinolinyl)-benzoic acid 380 Pro X₁ X₂ Leu Asp X₁ is 2-(5-quinolinyl)-phenylalanine-reduced Thr X₂ is beta-homolysine 381 Lys X Leu Asp Thr X is N-methyl beta-homolysine 

The invention claimed is:
 1. A homodimer of two moieties covalently linked together by an amide bond to a diphenic acid residue, the moieties both being of formula (I):

wherein R¹ is H; R² is H; R³ is CH₃; R⁴ is H; R⁵ is an amide of the formula —C(O)NR**R***, wherein R** is H and R*** is tert-butyl; R⁶ along with R⁷ together form the pyrrolidine ring of Pro; R⁸ is H; n is 1; wherein Z is an amino terminus of an amino acid; —C═O— adjacent L is the carboxy terminus of an amino acid; and L along with Z and —C═O— is a peptide having the following formula: X^(y)—X^(z)—X¹—X²—X³ wherein X^(y) is 3-aminomethyl-4-[4-(1-piperazinyl)-phenyl]-benzoic acid, in which the piperazinyl amine nitrogen at the 4-position of the piperazinyl ring is covalently bound via the amide bond to the diphenic acid residue to form the homodimer; X^(z) is absent; X¹ is Leucine; X² is Asp; and X³ is Thr; or a pharmaceutically acceptable salt thereof.
 2. A pharmaceutical composition comprising the homodimer of claim 1 along with a pharmaceutically acceptable carrier.
 3. The pharmaceutical composition of claim 2, formulated for oral delivery, topical delivery or parenteral delivery. 